Salts and polymorphs of 8-fluoro-2-{4-[(methylamino)methyl]phenyl}-1,3,4,5-tetrahydro-6h-azepino[5,4,3-cd]indol-6-one

ABSTRACT

The present invention relates to novel polymorphic forms of 8-fluoro-2-{4-[(methylamino)methyl]phenyl}-1,3,4,5-tetrahydro-6H-azepino[5,4,3-cd]indol-6-one, and to processes for their preparation. Such polymorphic forms may be a component of a pharmaceutical composition and may be used to treat a mammalian disease condition mediated by poly(ADP-ribose) polymerase activity including the disease condition such as cancer.

RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.14/698,463, filed Apr. 28, 2015, entitled “Salts and Polymorphs of8-Fluoro-2-{4-[(Methylamino)Methyl]Phenyl}-1,3,4,5-Tetrahydro-6H-Azepino[5,4,3-cd]Indol-6-One”.U.S. application Ser. No. 14/698,463 is a continuation of U.S.application Ser. No. 14/272,589, filed May 8, 2014, now U.S. Pat. No.9,045,487. U.S. application Ser. No. 14/272,589 is a continuation ofU.S. application Ser. No. 13/522,549, filed Jul. 17, 2012, now U.S. Pat.No. 8,754,072 which is a 35 U.S.C. § 371 national phase application ofInternational Application Serial No. PCT/IB2011/050571 (WO2011/098971A1), filed Feb. 10, 2011. International Application SerialNo. PCT/IB2011/050571 claims the benefit of U.S. Patent Application No.61/304,277, filed Feb. 12, 2010. Each of these applications isincorporated herein by reference in their entirety.

FIELD

The present invention relates to novel polymorphic salts of8-fluoro-2-{4-[(methylamino)methyl]phenyl}-1,3,4,5-tetrahydro-6H-azepino[5,4,3-cd]indol-6-one,and to methods for their preparation. The invention is also directed topharmaceutical compositions containing at least one polymorphic form andto the therapeutic and/or prophylactic use of such polymorphic forms andcompositions.

BACKGROUND

The compound8-fluoro-2-{4-[(methylamino)methyl]phenyl}-1,3,4,5-tetrahydro-6H-azepino[5,4,3-cd]indol-6-one(“Compound 1”)

is a small molecule inhibitor of poly(ADP-ribose) polymerase (PARP).Compound 1, and methods of making it, are described in U.S. Pat. Nos.6,495,541; 6,977,298; 7,429,578 and 7,323,562. Certain salts andpolymorphs thereof, of Compound 1, are disclosed in U.S. Pat. No.7,268,126 and in International Patent Publication No. WO 04/087713.Other publications describing Compound 1 and uses thereof include U.S.Patent Application Publication No. 2006-0074073, and U.S. Pat. Nos.7,351,701 and 7,531,530.

PARP is a family of nuclear enzymes responsible for ADP-ribosylation (apost-translational protein modification) in whichpoly(ADP-ribosyl)transferases transfer the ADP-ribose moiety from NAD⁺onto specific amino acid side chains on nuclear target proteins such ashistones and DNA repair enzymes and/or onto previously attachedADP-ribose units. In humans the PARP family encompasses 17 enzymes ofwhich PARP-1 is the best-characterized (Otto H, Reche P A, Bazan F etal, In silico characterization of the family of PARP-likepoly(ADP-ribosyl)transferases (pARTs), BMC Genomics 2005; 6:139).Pharmacology studies have shown that Compound 1 is an inhibitor ofPARP-1 (Ki=1.4 nM) and PARP-2 (Ki=0.17 nM).

PARP-1 is involved in DNA homeostasis through binding to DNA breaks andattracting DNA repair proteins to the site of DNA damage. PARP-1 throughthe addition of ADP-ribose units on target proteins provides theenergetic resources necessary for chromatin relaxation and the DNArepair process. These actions promote and facilitate DNA repair.Depending on the extent of DNA damage PARP-1 activation and subsequentpoly(ADP-ribosyl)ation mediate the repair of damaged DNA or induce celldeath. When DNA damage is moderate, PARP-1 plays a significant role inthe DNA repair process. Conversely, in the event of massive DNA damage,excessive activation of PARP-1 depletes the cellular ATP pool, whichultimately leads to cell mortality by necrosis (Tentori L, Portarena I,Graziani G, Potential applications of poly(ADP-ribose) polymerase (PARP)inhibitors, Pharmacol Res 2002; 45:73-85).

In cancer therapy, many useful drugs as well as ionizing radiation exerttheir therapeutic effect through DNA damage. Enzyme-mediated repair ofsingle- or double-strand DNA breaks is a potential mechanism ofresistance to radiotherapy or cytotoxic drugs whose mechanism of actiondepends on DNA damage. Inhibition of DNA repair pathway enzymes is thusa strategy for the potentiation of anticancer agents. Inhibition ofPARP-1 has shown to potentiate the activity of DNA-damaging agents andionizing radiation in vivo and in vitro. Accordingly, PARP has beenidentified as a therapeutic target for cancer therapy in combinationwith DNA damaging agents. (Tentori L, Leonetti C, Scarsella M, et al,Systemic administration of GPA 15427, a novel poly(ADP-ribose)polymerase-1 inhibitor, increases the antitumor activity of temozolomideagainst intracranial melanoma, glioma, lymphoma, Clin Cancer Res 2003;9:5370-9. Satoh M S, Poirier G G, Lindahl T, NAD(+)-dependent repair ofdamaged DNA by human cell extracts, J Biol Chem 1993; 268:5480-7.)

In addition to the potential role as chemopotentiator or radiosensitizeragents, more recent evidence has emerged of sensitivity of cell lines,homozygous for either the BRCA1 or BRCA2 mutation, to a PARP inhibitoralone. (Bryant H E, Schultz N, Thomas H D, et al, Specific killing ofBRCA-2 deficient tumors with inhibitors of poly(ADP-ribose) polymerase,Nature 2005; 434:913-7. Farmer H, McCabe N, Lord C J, et al, Targetingthe DNA repair defect in BRCA mutant cells as a therapeutic strategy,Nature 2005; 434:917-21.) Preliminary clinical data from a Phase I studywith a single-agent PARP inhibitor has recently been published (Yap T A,Boss D S, Fong M, et al, First in human phase I pharmacokinetic (PK) andpharmacodynamic (PD) study of KU-0059436 (Ku), a small moleculeinhibitor of poly ADP-ribose polymerase (PARP) in cancer patients (p)including BRCA 1/2 mutation carriers, (J Clin Oncol 2007; 25 (SupplementJune 20):3529).

It is desirable to have crystalline salts and polymorphic forms thereofthat possess properties amenable to reliable formulation andmanufacture.

SUMMARY OF THE INVENTION

Some embodiments disclosed herein provide a maleate salt of8-fluoro-2-{4-[(methylamino)methyl]phenyl}-1,3,4,5-tetrahydro-6H-azepino[5,4,3-cd]indol-6-one.In some embodiments, the maleate salt is crystalline. In someembodiments, the maleate salt is a crystalline anhydrous salt.

In some embodiments, the maleate salt has a powder X-ray diffractionpattern comprising one or more or two or more peaks at diffractionangles (2θ) selected from the group consisting of 6.0±0.2, 20.3±0.2, and21.7±0.2. In some embodiments, said powder X-ray diffraction pattern isobtained using copper K-alpha₁ X-rays at a wavelength of 1.5406Angstroms. In some embodiments, the maleate salt has a powder X-raydiffraction pattern comprising peaks at diffraction angles (20) of6.0±0.2, 20.3±0.2, and 21.7±0.2, wherein said powder X-ray diffractionpattern is obtained using copper K-alpha₁ X-rays at a wavelength of1.5406 Ångstroms. In further embodiments, the salt has a powder X-raydiffraction pattern comprising peaks at diffraction angles (2θ)essentially the same as shown in FIG. 1. In additional embodiments, thesalt has a differential scanning calorimetry thermogram essentially thesame as shown in FIG. 2. In some embodiments, the salt is asubstantially pure polymorph of maleate polymorph Form A.

In some embodiments, the maleate salt has a powder X-ray diffractionpattern comprising one or more or two or more peaks at diffractionangles (2θ) selected from the group consisting of 7.5±0.2, 11.3±0.2, and24.3±0.2. In some embodiments, said powder X-ray diffraction pattern isobtained using copper K-alpha₁ X-rays at a wavelength of 1.5406Ångstroms. In some embodiments, the maleate salt has a powder X-raydiffraction pattern comprising peaks at diffraction angles (2θ) of7.5±0.2, 11.3±0.2, and 24.3±0.2, wherein said powder X-ray diffractionpattern is obtained using copper K-alpha₁ X-rays at a wavelength of1.5406 Ångstroms. In further embodiments, the maleate salt has a powderX-ray diffraction pattern comprising peaks at diffraction angles (2θ)essentially the same as shown in FIG. 3 or FIG. 4. In some embodiments,the maleate salt has a solid state NMR spectrum comprising one or moreor two or more ¹³C chemical shifts selected from the group consisting of171.3±0.2, 112.4±0.2, and 43.8±0.2 ppm. In some embodiments, the maleatesalt has a solid state NMR spectrum comprising ¹³C chemical shifts at171.3±0.2, 112.4±0.2, and 43.8±0.2 ppm. In further embodiments, themaleate salt has a solid state NMR spectrum comprising ¹³C chemicalshifts at positions essentially the same as shown in FIG. 5. In someembodiments, the maleate salt has a solid state NMR spectrum comprisinga ¹⁹F chemical shift at −123.1±0.2 ppm. In further embodiments, themaleate salt has a solid state NMR spectrum comprising ¹⁹F chemicalshifts at positions essentially the same as shown in FIG. 6. In someembodiments, the maleate salt has a powder X-ray diffraction patterncomprising: one or more or two or more or three peaks at diffractionangles (2θ) selected from the group consisting of 7.5±0.2, 11.3±0.2, and24.3±0.2 obtained using copper K-alpha₁ X-rays at a wavelength of 1.5406Ångstroms; and: 1) a solid state NMR spectrum comprising one or more ortwo or more or three ¹³C chemical shifts selected from the groupconsisting of 171.3±0.2, 112.4±0.2, and 43.8±0.2 ppm; and/or 2) a solidstate NMR spectrum comprising a ¹⁹F chemical shift at −123.1±0.2 ppm. Inadditional embodiments, the salt has a differential scanning calorimetrythermogram essentially the same as shown in FIG. 7. In additionalembodiments, the salt has a dynamic vapor sorption isotherm essentiallythe same as shown in FIG. 8. In some embodiments, the maleate salt hasone or more FT-IR spectral peaks as shown in Table 6. In someembodiments, the maleate salt has one or more FT-Raman spectral peaks asshown in Table 7. In some embodiments, the maleate salt is asubstantially pure polymorph of maleate polymorph Form B. Someembodiments provide for a mixture of maleate polymorph Form A andmaleate polymorph Form B.

Additional embodiments provide a pharmaceutical composition comprising amaleate salt (e.g., maleate polymorph Form A or maleate polymorph Form Bor a mixture thereof). In some embodiments, the pharmaceuticalcomposition comprises a solid dosage form (e.g., a tablet). In someembodiments, the pharmaceutical composition comprises approximately10%-25% of the maleate salt, approximately 45%-60% microcrystallinecellulose, approximately 20%-35% dicalciaum phosphate anhydrous,approximately 0.1%-5% sodium starch glycolate (type A), andapproximately 0.1%-5% magnesium stearate. In some embodiments, thepharmaceutical composition comprises approximately 17.18% of the maleatesalt, approximately 52.55% microcrystalline cellulose, approximately26.27% dicalciaum phosphate anhydrous, approximately 3% sodium starchglycolate (type A), and approximately 1% magnesium stearate. Someembodiments provide a method of treating a mammalian disease conditionmediated by poly(ADP-ribose) polymerase activity, the method comprisingadministering to a mammal in need thereof a therapeutically effectiveamount of a pharmaceutical composition comprising a maleate salt (e.g.,maleate polymorph Form A or maleate polymorph Form B or a mixturethereof). Some embodiments provide a method of treating cancer in amammal, the method comprising administering to the mammal atherapeutically effective amount of a pharmaceutical compositioncomprising a maleate salt (e.g., maleate polymorph Form A or maleatepolymorph Form B or a mixture thereof).

Some embodiments disclosed herein relate to a camsylate salt of8-fluoro-2-{4-[(methylamino)methyl]phenyl}-1,3,4,5-tetrahydro-6H-azepino[5,4,3-cd]indol-6-one.In some embodiments, the camsylate salt is crystalline. In someembodiments, the camsylate salt is a crystalline anhydrous salt. In someembodiments, the camsylate is S-camsylate. In other embodiments, thecamsylate is R-camsylate.

In some embodiments, the camsylate salt has a powder X-ray diffractionpattern comprising one or more or two or more or three or more or fouror more peaks at diffraction angles (2θ) selected from the groupconsisting of 6.0±0.2, 12.2±0.2, 12.7±0.2, 14.8±0.2 16.7±0.2, and22.4±0.2. In some embodiments, the camsylate salt has a powder X-raydiffraction pattern comprising one or more or two or more or three peaksat diffraction angles (2θ) selected from the group consisting of12.2±0.2, 14.8±0.2, and 22.4±0.2. In some embodiments, the powder X-raydiffraction pattern is obtained using copper K-alpha₁ X-rays at awavelength of 1.5406 Ångstroms. In further embodiments, the camsylatesalt has a powder X-ray diffraction pattern comprising peaks atdiffraction angles (2θ) essentially the same as shown in FIG. 9 or 10.In some embodiments, the camsylate salt has a solid state NMR spectrumcomprising one or more or two or more ¹³C chemical shifts selected fromthe group consisting of 213.4±0.2, 171.8±0.2, and 17.3±0.2 ppm. In someembodiments, the camsylate salt has a solid state NMR spectrumcomprising ¹³C chemical shifts at 213.4±0.2, 171.8±0.2, and 17.3±0.2ppm. In further embodiments, the camsylate salt has a solid state NMRspectrum comprising ¹³C chemical shifts at positions essentially thesame as shown in FIG. 11. In some embodiments, the camsylate salt has asolid state NMR spectrum comprising one or more ¹⁹F chemical shiftsselected from the group consisting of −118.9±0.2 and −119.7 ppm±0.2. Insome embodiments, the camsylate salt has a solid state NMR spectrumcomprising ¹⁹F chemical shifts at −118.9±0.2 and −119.7 ppm±0.2. Infurther embodiments, the camsylate salt has a solid state NMR spectrumcomprising ¹⁹F chemical shifts at positions essentially the same asshown in FIG. 12. In some embodiments, the camsylate salt has a powderX-ray diffraction pattern comprising one or more or two or more or threeor more or four or more or five peaks at diffraction angles (2θ)selected from the group consisting of 6.0±0.2, 12.2±0.2, 12.7±0.2,14.8±0.2 16.7±0.2, and 22.4±0.2 obtained using copper K-alpha₁ X-rays ata wavelength of 1.5406 Ångstroms; and 1) a solid state NMR spectrumcomprising one or more or two or more or three ¹³C chemical shiftsselected from the group consisting of 213.4±0.2, 171.8±0.2, and 17.3±0.2ppm; and/or 2) a solid state NMR spectrum comprising one or more or two¹⁹F chemical shifts selected from the group consisting of −118.9±0.2 and−119.7 ppm±0.2. In additional embodiments, the salt has a differentialscanning calorimetry thermogram essentially the same as shown in FIG.13. In additional embodiments, the salt has a dynamic vapor sorptionisotherm essentially the same as shown in FIG. 14. In some embodiments,the camsylate salt has one or more FT-IR spectral peaks as shown inTable 12. In some embodiments, the camsylate salt has one or moreFT-Raman spectral peaks as shown in Table 13. In some embodiments, thesalt is a substantially pure polymorph of S-camsylate polymorph Form A.

In some embodiments, the camsylate salt has a powder X-ray diffractionpattern comprising peaks at diffraction angles (2θ) essentially the sameas shown in FIG. 15. In some embodiments, the salt is a substantiallypure polymorph of S-camsylate polymorph Form B. Some embodiments providefor a mixture of S-camsylate polymorph Form A and S-camsylate polymorphForm B.

In some embodiments, the camsylate salt has a powder X-ray diffractionpattern comprising peaks at diffraction angles (2θ) essentially the sameas shown in FIG. 18. In some embodiments, the camsylate salt has apowder X-ray diffraction pattern comprising one or more or two or moreor three peaks at diffraction angles (2θ) selected from the groupconsisting of 15.0±0.2, 21.8±0.2, and 24.7±0.2. In some embodiments, thecamsylate salt has a solid state NMR spectrum comprising one or more ¹³Cchemical shifts as shown in Table 16. In some embodiments, the camsylatesalt has one or more ¹⁹F chemical shifts as shown in Table 17. In someembodiments, the camsylate salt has a solid state NMR spectrumcomprising two or more ¹³C chemical shifts selected from the groupconsisting of 211.7±0.2, 132.5±0.2, and 19.4±0.2 ppm. In someembodiments, the camsylate salt has a solid state NMR spectrumcomprising ¹³C chemical shifts at 211.7±0.2, 132.5±0.2, and 19.4±0.2ppm. In some embodiments, the camsylate salt has a solid state NMRspectrum comprising a ¹⁹F chemical shift at −118.5±0.2. In someembodiments, the camsylate salt has one or more FT-IR spectral peaks asshown in Table 18. In some embodiments, the camsylate salt has one ormore FT-Raman spectral peaks as shown in Table 19. In some embodiments,the salt is a substantially pure polymorph of S-camsylate polymorph FormC. Some embodiments provide for a mixture of two or more of S-camsylatepolymorph Form A, S-camsylate polymorph Form B and S-camsylate polymorphForm C.

In some embodiments, the salt is a substantially pure polymorph ofR-camsylate polymorph Form A. Further embodiments provide additionalcamsylate salts. The salts can have various R:S ratios of camphorsulfonic acid, e.g., a 1R:1S-camsylate salt, a 1R:9S-camsylate salt, a1R:3S-camsylate salt, and a 1R:7S-camsylate salt.

Further embodiments provide for an amorphous form of the S-camsylatesalt of Compound 1.

Additional embodiments provide a pharmaceutical composition comprisingan camsylate salt described herein (e.g., S-camsylate polymorph Form A,S-camsylate polymorph Form B, S-camsylate polymorph Form C, R-camsylatepolymorph Form A or a mixture thereof). In some embodiments, thepharmaceutical composition comprises a solid dosage form (e.g., atablet). In some embodiments, the pharmaceutical composition comprisesapproximately 10%-25% of the camsylate salt, approximately 45%-60%microcrystalline cellulose, approximately 20%-35% dicalciaum phosphateanhydrous, approximately 0.1%-5% sodium starch glycolate (type A), andapproximately 0.1%-5% magnesium stearate. In some embodiments, thepharmaceutical composition comprises approximately 17.18% of thecamsylate salt, approximately 52.55% microcrystalline cellulose,approximately 26.27% dicalciaum phosphate anhydrous, approximately 3%sodium starch glycolate (type A), and approximately 1% magnesiumstearate. Some embodiments provide a method of treating a mammaliandisease condition mediated by poly(ADP-ribose) polymerase activity, themethod comprising administering to a mammal in need thereof atherapeutically effective amount of a pharmaceutical compositioncomprising an camsylate salt described herein (e.g., S-camsylatepolymorph Form A, S-camsylate polymorph Form B, S-camsylate polymorphForm C, R-camsylate polymorph Form A or a mixture thereof). Someembodiments provide a method of treating cancer in a mammal, the methodcomprising administering to the mammal a therapeutically effectiveamount of a pharmaceutical composition comprising an S-camsylate saltdescribed herein (e.g., S-camsylate polymorph Form A, S-camsylatepolymorph Form B, S-camsylate polymorph Form C, R-camsylate polymorphForm A or a mixture thereof).

Further embodiments provide a pharmaceutical composition comprising twoor more polymporph forms or salts described herein.

Additional embodiments provide methods of treating a mammalian diseasecondition mediated by poly(ADP-ribose) polymerase activity, the methodcomprising administering to a mammal in need thereof a therapeuticallyeffective amount of a pharmaceutical composition described herein incombination with a therapeutically effective amount of one or moresubstances, such as anti-tumor agents, anti-angiogenesis agents, signaltransduction inhibitors, and antiproliferative agents, mitoticinhibitors, alkylating agents, anti-metabolites, intercalatingantibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes,topoisomerase inhibitors, biological response modifiers, antibodies,cytotoxics, anti-hormones, and anti-androgens. Some embodiments providea method of treating cancer in a mammal, the method comprisingadministering to the mammal a therapeutically effective amount of apharmaceutical composition described herein in combination with atherapeutically effective amount of one or more substances, such asanti-tumor agents, anti-angiogenesis agents, signal transductioninhibitors, and antiproliferative agents, mitotic inhibitors, alkylatingagents, anti-metabolites, intercalating antibiotics, growth factorinhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors,biological response modifiers, antibodies, cytotoxics, anti-hormones,and anti-androgens.

Definitions

As used herein, the term “Compound 1” refers to the chemical compound8-fluoro-2-{4-[(methylamino)methyl]phenyl}-1,3,4,5-tetrahydro-6H-azepino[5,4,3-cd]indol-6-one,also represented by the structural formula:

The term “active agent” or “active ingredient” refers to a polymorphicform of Compound 1, or to a solid form that comprises two or morepolymorphic forms or amorphous form of Compound 1.

As used herein, the term “substantially pure” with reference to aparticular polymorphic form (or to a mixture of two or more polymorphicforms) of Compound 1 indicates the polymorphic form (or a mixture)includes less than 10%, preferably less than 5%, preferably less than3%, preferably less than 1% by weight of impurities, including otherpolymorphic forms of Compound 1. Such purity may be determined, forexample, by powder X-ray diffraction.

As used herein, the term “polymorph” refers to different crystallineforms of the same compound and other solid state molecular formsincluding pseudo-polymorphs, such as hydrates (e.g., bound water presentin the crystalline structure) and solvates (e.g., bound solvents otherthan water) of the same compound. Different crystalline polymorphs havedifferent crystal structures due to a different packing of the moleculesin the lattice. This results in a different crystal symmetry and/or unitcell parameters which directly influences its physical properties suchthe X-ray diffraction characteristics of crystals or powders. Adifferent polymorph, for example, will in general diffract at adifferent set of angles and will give different values for theintensities. Therefore X-ray powder diffraction can be used to identifydifferent polymorphs, or a solid form that comprises more than onepolymorph, in a reproducible and reliable way (S. Byrn et al,Pharmaceutical Solids: A Strategic Approach to RegulatoryConsiderations, Pharmaceutical research, Vol. 12, No. 7, p. 945-954,1995; J. K. Haleblian and W. McCrone, Pharmacetical Applications ofPolymorphism, Journal of Pharmaceutical Sciences, Vol. 58, No. 8, p.911-929, 1969). Crystalline polymorphic forms are of interest to thepharmaceutical industry and especially to those involved in thedevelopment of suitable dosage forms. If the polymorphic form is notheld constant during clinical or stability studies, the exact dosageform used or studied may not be comparable from one lot to another. Itis also desirable to have processes for producing a compound with theselected polymorphic form in high purity when the compound is used inclinical studies or commercial products since impurities present mayproduce undesired toxicological effects. Certain polymorphic forms mayexhibit enhanced thermodynamic stability or may be more readilymanufactured in high purity in large quantities, and thus are moresuitable for inclusion in pharmaceutical formulations. Certainpolymorphs may display other advantageous physical properties such aslack of hygroscopic tendencies, improved solubility, and enhanced ratesof dissolution due to different lattice energies.

The term “powder X-ray diffraction pattern” or “PXRD pattern” refers tothe experimentally observed diffractogram or parameters derivedtherefrom. Powder X-ray diffraction patterns are typically characterizedby peak position (abscissa) and peak intensities (ordinate). The term“peak intensities” refers to relative signal intensities within a givenX-ray diffraction pattern. Factors which can affect the relative peakintensities are sample thickness and preferred orientation (i.e., thecrystalline particles are not distributed randomly). The term “peakpositions” as used herein refers to X-ray reflection positions asmeasured and observed in powder X-ray diffraction experiments. Peakpositions are directly related to the dimensions of the unit cell. Thepeaks, identified by their respective peak positions, have beenextracted from the diffraction patterns for the various polymorphicforms of salts of Compound 1.

The term “2 theta value” or “26” refers to the peak position in degreesbased on the experimental setup of the X-ray diffraction experiment andis a common abscissa unit in diffraction patterns. In general, theexperimental setup requires that if a reflection is diffracted when theincoming beam forms an angle theta (θ) with a certain lattice plane, thereflected beam is recorded at an angle 2 theta (2θ). It should beunderstood that reference herein to specific 2θ values for a specificpolymorphic form is intended to mean the 2θ values (in degrees) asmeasured using the X-ray diffraction experimental conditions asdescribed herein.

The term “amorphous” refers to any solid substance which (i) lacks orderin three dimensions, or (ii) exhibits order in less than threedimensions, order only over short distances (e.g., less than 10 Å), orboth. Thus, amorphous substances include partially crystalline materialsand crystalline mesophases with, e.g. one- or two-dimensionaltranslational order (liquid crystals), orientational disorder(orientationally disordered crystals), or conformational disorder(conformationally disordered crystals). Amorphous solids may becharacterized by known techniques, including powder X-ray powderdiffraction (PXRD) crystallography, solid state nuclear magnet resonance(ssNMR) spectroscopy, differential scanning calorimetry (DSC), or somecombination of these techniques. Amorphous solids give diffuse PXRDpatterns, typically comprised of one or two broad peaks (i.e., peakshaving base widths of about 5° 2θ or greater).

The term “crystalline” refers to any solid substance exhibitingthree-dimensional order, which in contrast to an amorphous solidsubstance, gives a distinctive PXRD pattern with sharply defined peaks.

The term “ambient temperature” refers to a temperature conditiontypically encountered in a laboratory setting. This includes theapproximate temperature range of about 20 to about 30° C.

The term “detectable amount” refers to an amount or amount per unitvolume that can be detected using conventional techniques, such as X-raypowder diffraction, differential scanning calorimetry, HPLC, FourierTransform Infrared Spectroscopy (FT-IR), Raman spectroscopy, and thelike.

The term “solvate” describes a molecular complex comprising the drugsubstance and a stoichiometric or non-stoichiometric amount of one ormore solvent molecules (e.g., ethanol). When the solvent is tightlybound to the drug the resulting complex will have a well-definedstoichiometry that is independent of humidity. When, however, thesolvent is weakly bound, as in channel solvates and hygroscopiccompounds, the solvent content will be dependent on humidity and dryingconditions. In such cases, the complex will often be non-stoichiometric.

The term “hydrate” describes a solvate comprising the drug substance anda stoichiometric or non-stoichiometric amount of water.

The term “relative humidity” refers to the ratio of the amount of watervapor in air at a given temperature to the maximum amount of water vaporthat can be held at that temperature and pressure, expressed as apercentage.

The term “relative intensity” refers to an intensity value derived froma sample X-ray diffraction pattern. The complete ordinate range scalefor a diffraction pattern is assigned a value of 100. A peak havingintensity falling between about 50% to about 100% on this scaleintensity is termed very strong (vs); a peak having intensity fallingbetween about 50% to about 25% is termed strong (s). Additional weakerpeaks are present in typical diffraction patterns and are alsocharacteristic of a given polymorph.

The term “slurry” refers to a solid substance suspended in a liquidmedium, typically water or an organic solvent.

The term “under vacuum” refers to typical pressures obtainable by alaboratory oil or oil-free diaphragm vacuum pump.

The term “pharmaceutical composition” refers to a composition comprisingone or more of the polymorphic forms of salts of Compound 1 describedherein, and other chemical components, such asphysiologically/pharmaceutically acceptable carriers, diluents, vehiclesand/or excipients. The purpose of a pharmaceutical composition is tofacilitate administration of a compound to an organism, such as a humanor other mammal.

The term “pharmaceutically acceptable” “carrier”, “diluent”, “vehicle”,or “excipient” refers to a material (or materials) that may be includedwith a particular pharmaceutical agent to form a pharmaceuticalcomposition, and may be solid or liquid. Exemplary solid carriers arelactose, sucrose, talc, gelatin, agar, pectin, acacia, magnesiumstearate, stearic acid and the like. Exemplary liquid carriers aresyrup, peanut oil, olive oil, water and the like. Similarly, the carrieror diluent may include time-delay or time-release material known in theart, such as glyceryl monostearate or glyceryl distearate alone or witha wax, ethylcellulose, hydroxypropylmethylcellulose, methylmethacrylateand the like.

The term “mediated by poly(ADP-ribose) polymerase (PARP) activity”refers to biological or molecular processes that are regulated,modulated, or inhibited by PARP activity. For certain applications,inhibition of PARP activity associated with cancer is preferred.Embodiments disclosed herein include methods of modulating or inhibitingPARP activity, for example in mammals, by administering polymorphic saltforms of Compound 1, or a solid form that comprises two or morepolymorphic salt forms of Compound 1. The activity or efficacy ofpolymorphic salt forms of Compound 1, or a solid form that comprises twoor more such forms, may be measured as described, for example, in U.S.Pat. No. 6,495,541 and U.S. Patent Application Publication No.2006-0074073, the disclosures of which are incorporated herein byreference in their entireties.

The term “treating”, as used herein, unless otherwise indicated, meansreversing, alleviating, inhibiting the progress of, or preventing thedisorder or condition to which such term applies, or one or moresymptoms of such disorder or condition. The term “treatment”, as usedherein, unless otherwise indicated, refers to the act of “treating” asdefined immediately above. For example, the terms “treat”, “treating”and “treatment” can refer to a method of alleviating or abrogating ahyperproliferative disorder (e.g., cancer) and/or one or more of itsattendant symptoms. With regard particularly to cancer, these terms canindicate that the life expectancy of an individual affected with acancer will be increased or that one or more of the symptoms of thedisease will be reduced.

An “effective amount” refers to the amount of an agent thatsignificantly inhibits proliferation and/or prevents de-differentiationof a eukaryotic cell, e.g., a mammalian, insect, plant or fungal cell,and is effective for the indicated utility, e.g., specific therapeutictreatment.

The term “therapeutically effective amount” refers to that amount of thecompound or polymorph being administered which can relieve to someextent one or more of the symptoms of the disorder being treated. Inreference to the treatment of cancer, a therapeutically effective amountrefers to that amount which, for example, has at least one of thefollowing effects:

-   -   (1) reducing the size of the tumor;    -   (2) inhibiting (that is, slowing to some extent, preferably        stopping) tumor metastasis;    -   (3) inhibiting to some extent (that is, slowing to some extent,        preferably stopping) tumor growth, and    -   (4) relieving to some extent (or, preferably, eliminating) one        or more symptoms associated with the cancer.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a powder X-ray diffraction (PXRD) pattern of a maleate saltof Compound 1, polymorph Form A, using CuKα radiation at 1.5406 Å.

FIG. 2 shows a differential scanning calorimetry (DSC) thermogram of amaleate salt of Compound 1, polymorph Form A.

FIG. 3 shows a simulated PXRD pattern of a maleate salt of Compound 1,polymorph Form B, using CuKα radiation at 1.5406 Å.

FIG. 4 shows an experimental PXRD pattern of a maleate salt of Compound1, polymorph Form B, using CuKα radiation at 1.5406 Å.

FIG. 5 shows a ¹³C solid state nuclear magnetic resonance (NMR) spectrumof a maleate salt of Compound 1, polymorph Form B.

FIG. 6 shows a ¹⁹F solid state NMR spectrum of a maleate salt ofCompound 1, polymorph Form B.

FIG. 7 shows a DSC thermogram of a maleate salt of Compound 1, polymorphForm B.

FIG. 8 shows a dynamic vapor sorption isotherm of a maleate salt ofCompound 1, polymorph Form B.

FIG. 9 shows a simulated PXRD pattern of an S-camsylate salt of Compound1, polymorph Form A, using CuKα radiation at 1.5406 Å.

FIG. 10 shows an experimental PXRD pattern of an S-camsylate salt ofCompound 1, polymorph Form A, using CuKα radiation at 1.5406 Å.

FIG. 11 shows a ¹³C solid state NMR spectrum of an S-camsylate salt ofCompound 1, polymorph Form A.

FIG. 12 shows a ¹⁹F solid state NMR spectrum of an S-camsylate salt ofCompound 1, polymorph Form A.

FIG. 13 shows a DSC thermogram of an S-camsylate salt of Compound 1,polymorph Form A.

FIG. 14 shows a dynamic vapor sorption isotherm of an S-camsylate saltof Compound 1, S-camsylate polymorph Form A.

FIG. 15 shows PXRD pattern of an S-camsylate salt of Compound 1,polymorph Form B, using CuKα radiation at 1.5406 Å.

FIG. 16 shows an experimental PXRD pattern of a formulated compositioncontaining the S-camsylate salt of Compound 1, polymorph Form A.

FIG. 17 shows a simulated PXRD pattern of a hydrochloride salttrihydrate of Compound 1, using CuKα radiation at 1.5406 Å.

FIG. 18 shows an experimental PXRD pattern of an S-camsylate salt ofCompound 1, polymorph Form C, using CuKα radiation at 1.5406 Å.

FIG. 19 shows an experimental PXRD pattern of a 1R:1S-camsylate salt,using CuKα radiation at 1.5406 Å.

FIG. 20 shows an experimental PXRD pattern of a 1R:9S-camsylate salt,using CuKα radiation at 1.5406 Å.

FIG. 21 shows an experimental PXRD pattern of a 1R:3S-camsylate salt,using CuKα radiation at 1.5406 Å.

FIG. 22 shows an experimental PXRD pattern of a 1R:7S-camsylate salt,using CuKα radiation at 1.5406 Å.

FIG. 23 shows an experimental PXRD pattern of an R-camsylate salt ofCompound 1, polymorph Form A, using CuKα radiation at 1.5406 Å.

FIG. 24 shows a DSC thermogram of an S-camsylate salt of Compound 1,polymorph Form C.

FIG. 25 shows a DSC thermogram of a 1R:1 S-camsylate salt.

FIG. 26 shows a DSC thermogram of a 1R:9S-camsylate salt.

FIG. 27 shows a DSC thermogram of an R-camsylate salt of Compound 1,polymorph Form A.

FIG. 28 shows a ¹³C solid state NMR spectrum of an S-camsylate salt ofCompound 1, polymorph Form C.

FIG. 29 shows a ¹⁹F solid state NMR spectrum of an S-camsylate salt ofCompound 1, polymorph Form C.

FIG. 30 shows a ¹³C solid state NMR spectrum of a 1R:1S-camsylate salt.

FIG. 31 shows a ¹⁹F solid state NMR spectrum of a 1R:1S-camsylate salt.

FIG. 32 shows a ¹³C solid state NMR spectrum of a 1R:9S-camsylate salt.

FIG. 33 shows a ¹⁹F solid state NMR spectrum of a 1R:9S-camsylate salt.

FIG. 34 shows an experimental PXRD pattern of an amorphous form of theS-camsylate salt of Compound 1.

FIG. 35 shows a ¹³C solid state NMR spectrum of an amorphous form of theS-camsylate salt of Compound 1.

FIG. 36 shows a ¹⁹F solid state NMR spectrum of an amorphous form of theS-camsylate salt of Compound 1.

FIG. 37 shows a Raman spectrum of an amorphous form of the S-camsylatesalt of Compound 1.

FIG. 38 shows a DSC thermogram of an amorphous form of the S-camsylatesalt of Compound 1.

DETAILED DESCRIPTION OF THE INVENTION

Several unique physical forms of Compound 1 have now been made. Compound1, and methods of making it, are described in U.S. Pat. Nos. 6,495,541;6,977,298; 7,429,578 and 7,323,562, which are herein incorporated byreference in their entireties. Certain salts and polymorphs thereof, ofCompound 1, are disclosed in U.S. Pat. No. 7,268,126 and inInternational Patent Publication No. WO 04/087713, which are hereinincorporated by reference in their entireties.

It has been found, as described herein, that Compound 1 can exist inmultiple crystalline salt forms, such as maleate salt forms andcamsylate salt forms. These forms may be used in a formulated productfor the treatment of a mammalian disease condition mediated bypoly(ADP-ribose) polymerase (PARP) activity, including cancer. Each formmay have advantages over the others in terms of properties such asbioavailability, stability, and manufacturability. Novel crystallinesalt forms of Compound 1 have been discovered which are likely to bemore suitable for bulk preparation and handling than other forms. Forexample, the phosphate salt of Compound 1, while particularly suitable,for example, for intravenous dosage forms, may be less suitable for asolid dosage form due to its susceptibility to hydration. Maleate andcamsylate salt forms described herein (e.g., maleate polymorph Form Band S-camsylate polymorph Form A) exist as physically stable forms andare not susceptible to hydration as compared to other salt forms ofCompound 1, making them particularly suitable in the preparation ofsolid dosage forms. In addition, maleate and camsylate salts describedherein can be isolated in fewer steps than other salt forms in thesynthetic process, allowing greater scope to control thecrystallization. A controlled crystallization can be used, for example,to provide API particles with properties that are advantageous to asolid dosage form, such as controlled particle size, crystallinity andcrystal shape. Also described herein are processes for the preparationof each polymorphic salt form of Compound 1, substantially free fromother polymorphic forms of Compound 1. Additionally, described hereinare pharmaceutical formulations comprising crystalline salts of Compound1 in different polymorphic forms, and methods of treatinghyperproliferative conditions by administering such pharmaceuticalformulations. Additionally, described herein are pharmaceuticalformulations comprising crystalline salts of Compound 1 in differentpolymorphic forms, and methods of treating a mammalian disease condition(e.g., cancer) mediated by poly(ADP-ribose) polymerase (PARP) activityby administering such pharmaceutical formulations.

I. Crystalline Salt Forms of Compound 1

Several crystalline forms of Compound 1 are described herein. Eachcrystalline salt form of Compound 1 can be characterized by one or moreof the following: powder X-ray diffraction pattern (e.g., X-raydiffraction peaks at various diffraction angles (2θ)); solid statenuclear magnetic resonance (NMR) spectral pattern; melting point onset(and onset of dehydration for hydrated forms) as illustrated byendotherms of a Differential Scanning Calorimetry (DSC) thermogram;hygroscopic properties as illustrated by Dynamic Vapor Sorptionmeasurements; FT-IR spectral diagram pattern; Raman spectral diagrampattern; aqueous solubility; light stability under InternationalConference on Harmonization (ICH) high intensity light conditions, andphysical and chemical storage stability according to methods known inthe art or described herein. For example, maleate polymorph Form A,maleate polymorph Form B, S-camsylate polymorph Form A, and S-camsylatepolymorph Form B and of Compound 1 were each characterized by thepositions and relative intensities of peaks in their powder X-raydiffraction patterns. The powder X-ray diffraction parameters differ foreach of the polymorphic forms of Compound 1. For example, maleatepolymorph Form A, maleate polymorph Form B, S-camsylate polymorph FormA, and S-camsylate polymorph Form B of Compound 1 can therefore bedistinguished from each other and from other polymorphic forms ofCompound 1 by using powder X-ray diffraction.

Powder X-ray diffraction patterns of the different polymorphic forms(e.g., maleate polymorph Form A, maleate polymorph Form B, S-camsylatepolymorph Form A, and S-camsylate polymorph Form B) of Compound 1 weredetermined according to procedures described in Examples 6-8 using CuKαradiation at 1.5406 Å. The peaks for the PXRD patterns obtained forMaleate polymorph Form A, Maleate polymorph Form B, S-camsylatepolymorph Form A, and S-camsylate polymorph Form B were selected usingBruker-AXS Ltd. Evaluation software with a threshold of 1 and a peakwidth of 0.3° 2-theta. With the exception of S-camsylate polymorph FormB, the data were collected at 21° C.

To perform an X-ray diffraction measurement on a Bragg-Brentanoinstrument like the Bruker system used for measurements reported herein,the sample is typically placed into a holder which has a cavity. Thesample powder is pressed by a glass slide or equivalent to ensure arandom surface and proper sample height. The sample holder is thenplaced into the instrument. The incident X-ray beam is directed at thesample, initially at a small angle relative to the plane of the holder,and then moved through an arc that continuously increases the anglebetween the incident beam and the plane of the holder. Measurementdifferences associated with such X-ray powder analyses can result from avariety of factors including: (a) errors in sample preparation (e.g.,sample height); (b) instrument errors (e.g., flat sample errors); (c)calibration errors; (d) operator errors (including those errors presentwhen determining the peak locations); and (e) the nature of the material(e.g., preferred orientation and transparency errors). Calibrationerrors and sample height errors often result in a shift of all the peaksin the same direction. Small differences in sample height when using aflat holder will lead to large displacements in PXRD peak positions. Asystematic study showed that, using a Shimadzu XRD-6000 in the typicalBragg-Brentano configuration, sample height difference of 1 mm led topeak shifts as high as 1 degree (2θ)□ (Chen et al., J Pharmaceutical andBiomedical Analysis 26:63 (2θ01)). These shifts can be identified fromthe X-ray diffractogram and can be eliminated by compensating for theshift (applying a systematic correction factor to all peak positionvalues) or recalibrating the instrument. It is possible to rectifymeasurements from the various machines by applying a systematiccorrection factor to bring the peak positions into agreement. Ingeneral, this correction factor will bring the measured peak positionsfrom the Bruker into agreement with the expected peak positions and maybe in the range of 0 to 0.2 degrees (2θ)□.

One of skill in the art will appreciate that the peak positions (2θ)will show some variability, typically as much as 0.1 to 0.2 degrees(2θ0), depending, for example, on the solvents being used and/or on theapparatus being used to measure the diffraction. Accordingly, where peakpositions (2θ) are reported, one of skill in the art will recognize thatsuch numbers are intended to encompass such variability. Furthermore,where the polymorphs of the present invention are described as having apowder X-ray diffraction pattern essentially the same as that shown in agiven figure, the term “essentially the same” is also intended toencompass such variability in diffraction peak positions. Further, oneskilled in the art will appreciate that relative peak intensities willshow inter-apparatus variability as well as variability due to thedegree of crystallinity, preferred orientation, prepared sample surface,the degree of purity of the sample being analyzed, and other factorsknown to those skilled in the art, and should be taken as qualitativemeasures only. The skilled person will also appreciate that measurementsusing a different wavelength will result in different shifts accordingto the Bragg equation—nλ=2d sin θ. Such further PXRD patterns generatedby use of alternative wavelengths are considered to be alternativerepresentations of the PXRD patterns of the crystalline materials ofembodiments described herein and as such are within the scope of thepresent embodiments.

The different polymorphs described herein can also be characterizedusing solid state NMR spectroscopy according to methods known in the artor described herein. For example, ¹³C solid state spectra and ¹⁹F solidstate spectra can be collected according to the procedures described inExamples 9-10. It should be noted that ¹³C or ¹⁹F chemical shiftsmeasured in solid state NMR will typically have a variability of up to0.2 ppm for well defined peaks, and even larger for broad lines.

Different crystalline salt forms of Compound 1 were also distinguishedusing differential scanning calorimetry (DSC) according to theprocedures described in the Examples. DSC measures the difference inheat energy uptake between a sample and an appropriate reference withincrease in temperature. For example, for the measurement of a solidpowder sample, the reference can be an empty sample pan of the type usedin preparation of the sample. DSC thermograms can be characterized byendotherms (indicating energy uptake) and also by exotherms (indicatingenergy release), typically as the sample is heated. Depending on severalfactors, the endotherms exhibited may vary by about 0.01-5° C. forcrystal polymorphs melting above or below the endotherms, such as thosedepicted in the appended figures. Factors responsible for such varianceinclude, for example, the rate of heating (e.g., the scan rate) at whichthe DSC analysis is conducted, the way the DSC onset temperature isdefined and determined, the calibration standard used, instrumentcalibration, the relative humidity and the chemical purity of thesample. For any given sample, the observed endotherms may also differfrom instrument to instrument; however, it will generally be within theranges described herein provided the instruments are calibratedsimilarly.

Different polymorphic forms of a compound may have different hygroscopicproperties. For example, salts of Compound 1 were characterized based ontheir hygroscopic properties using dynamic vapor sorption measurementsaccording to procedures described in Example 12.

In some embodiments, the solid forms may also comprise more than onepolymorphic form. One of skill in the art will also recognize thatcrystalline forms of a given compound can exist in substantially pureforms of a single polymorph, but can also exist in a crystalline formthat comprises a mixture of two or more different polymorphs oramorphous forms. Where a solid form comprises two or more polymorphs,the X-ray diffraction pattern will typically have peaks characteristicof each of the individual polymorphs. For example, a solid form thatcomprises two polymorphs will typically have a powder X-ray diffractionpattern that is a convolution of the two X-ray diffraction patterns thatcorrespond to the substantially pure polymorphic forms. For example, asolid form of Compound 1 or a salt thereof can contain a first andsecond polymorphic form where the solid form contains at least 10% byweight of the first polymorph. In a further example, the solid form cancontain at least 20% by weight of the first polymorph. Even furtherexamples contain at least 30%, at least 40%, or at least 50% by weightof the first polymorph. One of skill in the art will recognize that manysuch combinations of several individual polymorphs and amorphous formsin varying amounts are possible.

Two polymorphic forms of the maleate salt of Compound 1 have beenidentified and characterized as indicated in FIGS. 1 to 8, and aredesignated as maleate polymorph Form A and maleate polymorph Form B. Inaddition, polymorphic forms of the camsylate salt of Compound 1 andvarious salts containing different R:S ratios of camphor sulfonic acidhave been identified and characterized as indicated in FIGS. 9 to 33,and are designated as S-camsylate polymorph Form A, S-camsylatepolymorph Form B, S-camsylate polymorph Form C, R-camsylate polymorphForm A, or the salt with the designated R:S ratio of camphor sulfonicacid. Furthermore, an amorphous form of the S-camsylate salt of Compound1 has been identified and characterized as indicated in FIGS. 34-38. Asused herein, the term “camsylate salt” refers to the S-camsylate salt,the R-camsylate salt, or salts with camphor sulfonic acid in particularR:S ratios. The polymorphs, pharmaceutical compositions including one ormore polymorphs, and methods of using the polymorphs and pharmaceuticalcompositions thereof are described in more detail in the followingsections and examples.

A. Maleate Salt of Compound 1, Polymorph Form A

The maleate salt of Compound 1, maleate polymorph Form A, can beproduced as described in Example 1.

Maleate polymorph Form A was characterized by the PXRD pattern shown inFIG. 1 and described in Example 7. The PXRD pattern of maleate polymorphForm A, expressed in terms of the degree (2θ) and relative intensitieswith a relative intensity of ≧15.0%, measured on a Bruker D5000diffractometer with CuKα radiation at 1.5406 Å, is also shown in Table1.

TABLE 1 Angle (Degree 2θ ± 0.2°) Relative Intensity (≥15.0%) 6.0 50.912.0 44.7 13.8 15.8 14.8 29.4 15.5 40.3 17.9 35.6 19.8 25.5 20.3 39.520.9 26.7 21.7 32.4 23.3 100.0 24.0 42.5 24.5 25.2 24.8 25.2 25.4 24.526.2 19.5 27.5 16.7 28.3 19.0 29.2 20.5 30.3 20.5 31.0 17.4 36.8 15.5

The DSC thermogram for maleate polymorph Form A, shown in FIG. 2 anddescribed in Example 11, indicates an endotherm onset at 220.36° C.

B. Maleate Salt of Compound 1, Maleate Polymorph Form B

The maleate salt of Compound 1, maleate polymorph Form B, can beproduced as described in Example 2, using ethanol in the syntheticscheme. The maleate salt of Compound 1, maleate polymorph Form B, canalso be produced as described in Example 3, using isopropyl alcohol inthe synthetic scheme.

Maleate polymorph Form B was characterized by the simulated PXRD patterncalculated from a single crystal structure, as shown in FIG. 3. Thesimulated PXRD pattern of maleate polymorph Form B, expressed in termsof the degree (2θ) and relative intensities with a relative intensity of≧5.0%, calculated from the single crystal structure of maleate Form Busing the “Reflex Powder Diffraction” module of Accelrys MS Modelling™[version 4.4], is also shown in Table 2. Pertinent simulation parametersincluded a wavelength of 1.5406 Å (Cu Kα) and a polarization factor of0.5.

TABLE 2 Angle (Degree 2θ) Relative Intensity (≥5.0%) 11.3 5.5 11.4 12.214.0 5.4 14.7 5.1 15.1 5.1 15.5 32.9 15.7 5.1 16.1 8.5 16.5 11.1 17.934.5 19.9 8.2 21.0 17.7 24.2 7.1 24.6 7.0 24.8 100.0 26.2 6.4 27.4 6.427.7 16.2

Maleate polymorph Form B was also characterized by measuring the PXRDpattern for a particular batch of maleate polymorph Form B. Thisexperimental PXRD pattern is shown in FIG. 4 and described in Example 6.The experimental PXRD pattern of maleate polymorph Form B, expressed interms of the degree (2θ) and relative intensities with a relativeintensity of ≧5.0%, measured on a Bruker-AXS Ltd., D4 diffractometerwith CuKα radiation at 1.5406 Å, is also shown in Table 3.

TABLE 3 Angle (Degree 2θ ± 0.2°) Relative Intensity (≥5.0%) 7.5 14.410.4 26.6 11.3 9.0 12.9 5.4 13.9 9.4 15.1 33.1 15.5 61.1 15.7 28.2 16.16.2 16.4 27.3 17.9 18.6 19.9 6.8 20.9 100.0 22.7 5.8 23.5 7.6 24.3 28.624.6 16.5 24.8 59.2 26.2 9.3 26.6 7.5 27.1 5.7 27.3 8.8 27.7 34.9 28.05.7 30.4 5.0 31.7 15.3 32.0 5.6 33.3 6.3 40.4 5.1

It can be seen that the peak positions for the simulated andexperimental PXRD patterns agree very well. Any difference in peakposition, relative intensity and width of the diffraction peaks can beattributed, for example, to inter-apparatus variability as well asvariability due to the degree of crystallinity, preferred orientation,prepared sample surface, the degree of purity of the sample beinganalyzed, and other factors known to those skilled in the art.

Maleate polymorph Form B of Compound 1 was also characterized by thesolid state NMR spectral pattern shown in FIG. 5, carried out on aBruker-Biospin 4 mm BL CPMAS probe positioned into a wide-boreBruker-Biospin DSX 500 MHz NMR spectrometer as described in Example 9.The ¹³C chemical shifts of maleate polymorph Form B of Compound 1 areshown in Table 4.

TABLE 4 ¹³C Chemical Shifts^(a) [±0.2 ppm] Intensity^(b) 171.3 11.7169.6 7.3 160.5 2.6 158.6 3.4 137.7 10.4 136.4 9.8 134.1 10.8 132.7 12.0130.9 11.6 128.7 10.6 125.7 5.4 124.2 3.9 112.4 7.8 109.6 7.8 102.3 7.652.2 8.7 43.8 8.7 32.3 11.7 29.9 8.9 ^(a)Referenced to external sampleof solid phase adamantane at 29.5 ppm. ^(b)Defined as peak heights.Intensities can vary depending on the actual setup of the CPMASexperimental parameters and the thermal history of the sample. CPMASintensities are not necessarily quantitative.

Maleate polymorph Form B of Compound 1 was also characterized by thesolid state NMR spectral pattern shown in FIG. 6, carried out on aBruker-Biospin 4 mm BL CPMAS probe positioned into a wide-boreBruker-Biospin DSX 500 MHz NMR spectrometer as described in Example 9.The ¹⁹F chemical shifts of maleate polymorph Form B of Compound 1 areshown in Table 5.

TABLE 5 ¹⁹F Chemical Shifts^(a) [±0.2 ppm] Intensity^(b) −123.1 12.0^(a)Referenced to external standard of trifluoroacetic acid (50% V/V inH₂O) at −76.54 ppm. ^(b)Defined as peak heights.

The DSC thermogram for maleate polymorph Form B, shown in FIG. 7,indicates an endotherm onset at 228.0° C. The dynamic vapor sorptionisotherm for maleate polymorph Form B is shown in FIG. 8. The dynamicvapor sorption isotherm indicates maleate polymorph Form B isnon-hygroscopic.

Maleate polymorph Form B of Compound 1 was also characterized by FourierTransform-Infrared Spectroscopy (FT-IR) as described in Example 25, andthe spectral peaks are shown in Table 6. Absorption band frequencies arelisted. (w: weak, m: medium, s: strong, vs: very strong). Experimentalerror is ±2 cm⁻¹ except for * error on peak position could beconsiderably larger.

TABLE 6 Wavenumber (cm⁻¹) 3179* w 2970w 2927w 2884w 2830w 2484w 1685w1594m 1576m 1509w 1457s 1444s 1417m 1389w 1368m 1353s 1347s 1332s 1315m1275w 1267w 1252w 1212w 1179w 1159w 1127s 1106m 1066m 1051m 1030m 1020m1013m  971m  954m  938w  916w  895w  886m  877w  866s  856m  841s  836s 788s  761s  741m  699w  679s  663m

Maleate polymorph Form B of Compound 1 was also characterized by FourierTransform-Raman Spectroscopy (FT-Raman) as described in Example 26, andthe spectral peaks are shown in Table 7. (w: weak, m: medium, s: strong,vs: very strong).

Experimental error is ±2 cm⁻¹.

TABLE 7 Wavenumber (cm⁻¹) 3237w 3060w 3031w 2972w 2948w 2929w 2887w2834w 2819w 2716w 2651w 2589w 2562w 2534w 1694w 1621vs 1585s 1563s 1511m1460s 1431w 1407w 1387w 1370m 1350s 1330m 1268w 1218w 1195w 1181w 1130w1069s 1033w 1003w  961w  940w  898w  883w  857w  846w  794w  744w  732w 702w  665w  647w  619w  557w  524w  503w  487w  464w  433w  414w  402w 381w  345w  318w  299w  257w  216w  166w  149w  126m  106m  72s

C. S-Camsylate Salt of Compound 1, S-Camsylate Polymorph Form A

The S-camsylate salt of Compound 1, S-camsylate polymorph Form A, can beproduced as described in Example 4, using tetrahydrofuran in thesynthetic scheme. The S-camsylate salt of Compound 1, S-camsylatepolymorph Form A, can also be produced as described in Example 5, usingisopropyl alcohol in the synthetic scheme.

S-camsylate polymorph Form A was characterized by the simulated PXRDpattern calculated from a single crystal structure, as shown in FIG. 9.The simulated PXRD pattern of S-camsylate polymorph Form A, expressed interms of the degree (2θ) and relative intensities with a relativeintensity of ≧15.0%, calculated from the single crystal structure ofcamsylate Form A using the “Reflex Powder Diffraction” module ofAccelrys MS Modelling™ [version 4.4], is also shown in Table 8.Pertinent simulation parameters included a wavelength of 1.5406 Å (CuKα) and a polarization factor of 0.5.

TABLE 8 Angle (Degree 2θ) Relative Intensity (≥15.0%) 3.0 21.1 6.1 68.212.2 51.7 12.7 100.0 13.4 65.7 13.8 27.3 14.3 54.7 14.8 39.2 15.9 27.516.1 37.7 16.7 23.6 18.2 29.3 18.3 19.0 18.4 40.1 18.9 16.2 19.0 18.819.5 31.8 20.5 50.3 21.0 55.7 21.1 28.0 22.4 27.6 22.7 18.9 23.0 31.024.0 35.0 25.4 26.3 25.7 92.8 28.4 15.8

S-camsylate polymorph Form A was also characterized by measuring thePXRD pattern for a particular batch of S-camsylate polymorph Form A.This experimental PXRD pattern is shown in FIG. 10. The experimentalPXRD pattern of S-camsylate polymorph Form A, expressed in terms of thedegree (2θ) and relative intensities with a relative intensity of≧10.0%, measured on a Bruker-AXS Ltd., D4 diffractometer with CuKαradiation at 1.5406 Å, is also shown in Table 9.

TABLE 9 Angle (Degree 2θ ± 0.2°) Relative Intensity (≥10.0%) 6.0 22.912.2 100.0 12.7 28.8 13.5 46.2 13.8 20.8 14.3 11.9 14.8 59.5 15.2 14.416.1 12.5 16.3 13.5 16.7 32.3 18.3 54.8 18.5 12.9 19.5 55.4 20.5 30.321.1 34.1 22.5 58.8 22.7 10.7 23.1 19.8 24.1 15.6 24.5 22.3 25.4 49.925.7 56.0 27.4 17.0 28.5 11.8 29.8 17.2 30.7 20.6 30.8 18.8 31.5 13.7

It can be seen that the peak positions for the simulated andexperimental PXRD patterns agree very well. Any difference in peakposition, relative intensity and width of the diffraction peaks can beattributed, for example, to inter-apparatus variability as well asvariability due to the degree of crystallinity, preferred orientation,prepared sample surface, the degree of purity of the sample beinganalyzed, and other factors known to those skilled in the art.

S-camsylate polymorph Form A of Compound 1 was also characterized by thesolid state NMR spectral pattern shown in FIG. 11, carried out on aBruker-Biospin 4 mm BL CPMAS probe positioned into a wide-boreBruker-Biospin DSX 500 MHz NMR spectrometer as described in Example 10.The ¹³C chemical shifts of S-camsylate polymorph Form A of Compound 1are shown in Table 10.

TABLE 10 ¹³C Chemical Shifts^(a) [±0.2 ppm] Intensity^(b) 214.7 4.3213.4 4.0 171.8 5.6 160.7 1.8 160.0 2.0 158.7 2.5 158.0 2.5 137.6 4.5137.2 4.5 134.9 4.1 134.0 4.2 132.2 12.0 128.8 5.8 127.2 11.0 125.8 4.2124.7 4.1 123.2 5.9 113.2 6.5 110.1 4.8 102.8 2.6 102.0 3.0 58.6 10.153.0 4.1 52.5 4.4 49.3 5.9 48.0 9.8 42.8 10.6 41.8 4.7 37.4 3.8 35.3 3.832.5 2.8 31.0 2.9 28.2 5.8 27.0 3.5 25.0 3.5 20.1 5.0 18.4 8.8 17.3 4.5^(a)Referenced to external sample of solid phase adamantane at 29.5 ppm.^(b)Defined as peak heights. Intensities can vary depending on theactual setup of the CPMAS experimental parameters and the thermalhistory of the sample. CPMAS intensities are not necessarilyquantitative.

The S-camsylate polymorph Form A of Compound 1 was also characterized bythe solid state NMR spectral pattern shown in FIG. 12, carried out on aBruker-Biospin 4 mm BL CPMAS probe positioned into a wide-boreBruker-Biospin DSX 500 MHz NMR spectrometer as described in Example 10.The ¹⁹F chemical shifts of S-camsylate polymorph Form A of Compound 1are shown in Table 11.

TABLE 11 ¹⁹F Chemical Shifts^(a) [±0.2 ppm] Intensity^(b) −118.9 12.0−119.7 11.7 ^(a)Referenced to external standard of trifluoroacetic acid(50% V/V in H₂O) at −76.54 ppm. ^(b)Defined as peak heights.

The DSC thermogram for S-camsylate polymorph Form A, shown in FIG. 13,indicates an endotherm onset at 303.2° C. The dynamic vapor sorptionisotherm for S-camsylate polymorph Form A is shown in FIG. 14. Thedynamic vapor sorption isotherm indicates S-camsylate polymorph Form Ais non-hygroscopic.

S-camsylate polymorph Form A of Compound 1 was also characterized byFourier Transform-Infrared Spectroscopy (FT-IR) as described in Example25, and the spectral peaks are shown in Table 12. Absorption bandfrequencies are listed. (w: weak, m: medium, s: strong, vs: verystrong). Experimental error is ±2 cm⁻¹ except for * error on peakposition could be considerably larger.

TABLE 12 Wavenumber (cm⁻¹) 3287m 3237m 3074w 2962m 2949w 2892w 2839w1743s 1637s 1615s 1581w 1510w 1474m 1451m 1415m 1366w 1348w 1315m 1289w1266m 1255m 1240m 1234m 1226m 1202s 1193s 1151s 1128s 1103s 1066m 1056w1030s 1015s  979w  967w  958w  936w  898w  870m  864m  848m  834m  811m 787s  753m  720m  706m  674m

S-camsylate polymorph Form A of Compound 1 was also characterized byFourier Transform-Raman Spectroscopy (FT-Raman) as described in Example26, and the spectral peaks are shown in Table 13. (w: weak, m: medium,s: strong, vs: very strong). Experimental error is ±2 cm⁻¹.

TABLE 13 Wavenumber (cm⁻¹) 3299w 3230w 3109w 3076w 3059w 3043w 3024w3000w 2968m 2942w 2922w 2895w 2843w 2820w 2777w 2736w 2554w 1746w 1617vs1581s 1554vs 1510m 1454vs 1434m 1419w 1408w 1369m 1348s 1324s 1270w1251w 1214w 1200w 1160w 1133w 1068s 1041w 1022w  939w  901w  859w  816w 726w  689w  645w  621w  585w  550w  516w  503w  430w  416w  401w  370w 350w  278w  261w  243w  219w  158m  137w  115m  84m  64s

D. S-Camsylate Salt of Compound 1, S-Camsylate Polymorph Form B

The S-camsylate salt of Compound 1, S-camsylate polymorph Form B, wascharacterized by the PXRD pattern shown in FIG. 15.

E. Hydrochloride Salt Trihydrate Polymorph of Compound 1

A hydrochloride salt trihydrate polymorph of Compound 1 wascharacterized by the simulated PXRD pattern calculated from a singlecrystal structure, as shown in FIG. 17, using CuKα radiation at 1.5406Å. The simulated PXRD pattern of the hydrochloride salt trihydratepolymorph, expressed in terms of the degree (2θ) and relativeintensities with a relative intensity of ≧15.0%, is also shown in Table14.

TABLE 14 Angle (Degree 2θ) Relative Intensity (≥15.0%) 6.2 55.1 11.056.5 11.2 56.7 11.6 23.1 14.9 17.6 15.2 31.5 15.9 35.3 16.2 40.9 17.045.4 18.4 37.9 18.7 28.9 19.4 42.1 19.7 20.3 20.3 55.1 20.7 35.7 21.139.6 21.5 35.1 21.8 20.5 22.9 18.3 23.4 50.5 24.5 100.0 25.1 35.4 25.343.4 26.1 76.9 27.1 38.0 27.6 24.6 28.0 28.1 28.3 43.0 28.6 22.0 28.930.7 29.2 23.2 29.6 27.9 30.1 19.9 30.4 29.0 30.6 27.2 31.1 16.3 31.920.8 32.2 30.3 32.8 24.6 34.1 16.0 34.4 19.5 34.7 19.5 35.3 17.3 36.217.4 36.5 15.7 36.8 24.3 37.2 18.9 37.7 17.6 38.0 23.8 38.6 20.7 38.818.6 39.7 17.9

F. S-camsylate Salt of Compound 1, S-camsylate Polymorph Form C

The S-camsylate salt of Compound 1, S-camsylate polymorph Form C, can beproduced as described in Example 16.

S-camsylate polymorph Form C was characterized by measuring the PXRDpattern for a particular batch of S-camsylate polymorph Form C. Thisexperimental PXRD pattern is shown in FIG. 18. The experimental PXRDpattern of S-camsylate polymorph Form C, expressed in terms of thedegree (2θ) and relative intensities with a relative intensity greaterthan 10.0%, measured on a Bruker-AXS Ltd., D4 diffractometer with CuKαradiation at 1.5406 Å, is also shown in Table 15.

TABLE 15 Angle (Degree 2θ ± 0.1°) Relative Intensity (>10.0%) 6.0 13.411.9 22.7 12.7 89.6 13.5 75.6 14.2 16.0 14.6 18.8 15.0 33.2 15.2 34.516.6 24.5 17.9 32.7 18.6 45.7 19.1 17.2 19.7 17.1 20.6 42.2 21.0 17.521.8 32.8 22.9 18.8 23.4 26.6 24.0 27.8 24.7 100.0 25.7 26.1 27.9 17.429.6 18.4 30.1 27.6 33.0 15.4

S-camsylate polymorph Form C of Compound 1 was also characterized by thesolid state NMR spectral pattern shown in FIG. 28, carried out on aBruker-Biospin 4 mm BL CPMAS probe positioned into a wide-boreBruker-Biospin DSX 500 MHz NMR spectrometer as described in Example 23.The ¹³C chemical shifts of S-camsylate polymorph Form A of Compound 1are shown in Table 16.

TABLE 16 ¹³C Chemical Shifts^(a) [ppm] Intensity^(b) 214.6 3.6 211.7 3.0171.6 6.2 159.9 2.6 158.0 3.5 137.3 6.5 135.0 3.8 133.9 4.2 132.5 12.0128.5 10.6 126.9 7.2 124.6 4.1 123.3 5.6 113.8 3.7 113.1 4.8 110.0 4.3103.4 1.9 100.8 2.0 58.2 8.0 52.8 4.9 48.1 10.6 42.9 9.8 42.2 9.1 36.16.4 31.5 4.4 28.3 3.9 27.3 3.7 24.8 5.8 19.4 5.4 18.1 3.6 ^(a)Referencedto external sample of solid phase adamantane at 29.5 ppm. ^(b)Defined aspeak heights. Intensities can vary depending on the actual setup of theCPMAS experimental parameters and the thermal history of the sample.CPMAS intensities are not necessarily quantitative.

The S-camsylate polymorph Form C of Compound 1 was also characterized bythe solid state NMR spectral pattern shown in FIG. 29, carried out on aBruker-Biospin 4 mm BL CPMAS probe positioned into a wide-boreBruker-Biospin DSX 500 MHz NMR spectrometer as described in Example 23.The ¹⁹F chemical shifts of S-camsylate polymorph Form A of Compound 1are shown in Table 17.

TABLE 17 ¹⁹F Chemical Shifts^(a) [ppm] Intensity^(b) −118.5 12.0^(a)Referenced to external standard of trifluoroacetic acid (50% V/V inH₂O) at −76.54 ppm. ^(b)Defined as peak heights.

The DSC thermogram for S-camsylate polymorph Form C, shown in FIG. 24,indicates an endotherm onset at 291.9° C.

S-camsylate polymorph Form C of Compound 1 was also characterized byFourier Transform-Infrared Spectroscopy (FT-IR) as described in Example25, and the spectral peaks are shown in Table 18. Absorption bandfrequencies are listed. (w: weak, m: medium, s: strong, vs: verystrong). Experimental error is ±2 cm⁻¹ except for * error on peakposition could be considerably larger.

TABLE 18 Wavenumber (cm⁻¹) 3284m 3074w 3024w 2962m 2912w 2891w 2839w2581w 1753m 1743m 1637m 1615s 1582w 1513w 1472m 1451s 1415m 1367w 1346m1324m 1315m 1261m 1240s 1204m 1192m 1175m 1153s 1131s 1106s 1067m 1030s1024s  965w  958w  937w  899w  871m  843m  810m  787s  752w  721w  706w 674m

S-camsylate polymorph Form C of Compound 1 was also characterized byFourier Transform-Raman Spectroscopy (FT-Raman) as described in Example26, and the spectral peaks are shown in Table 19. (w: weak, m: medium,s: strong, vs: very strong). Experimental error is ±2 cm⁻¹.

TABLE 19 Wavenumber (cm⁻¹) 3291w* 3229w 3074w 3057w 3029w 2967w 2946w2915w 2892w 2844w 2819w 2777w 2732w 2554w 1755w 1745w 1617vs 1579s1555vs 1511w 1454vs 1408w 1369m 1348m 1324m 1269w 1250w 1217w 1204w1164w 1134w 1069s 1041w 1022w  960w  939w  902w  859w  815w  791w  726w 708w  683w  646w  636w  616w  582w  549w  504w  485w  430w  413w  370w 350w  275w  262w  242w  222w  160w  114m  89m  61m

G. R-Camsylate and S-Camsylate Salts

Various camsylate salts with different R:S ratios of camphor sulfonicacid were produced and characterized. The 1R:1S-camsylate salt, the1R:9S-camsylate salt, the 1R:3S-camsylate salt, and the 1R:7S-camsylatesalt can be produced as described in Examples 17-20.

The 1R:1S-camsylate salt, the 1R:9S-camsylate salt, the 1R:3S-camsylatesalt, and the 1R:7S-camsylate salt were characterized by measuring thePXRD pattern for a particular batch of each salt. These experimentalPXRD patterns are shown in FIGS. 19-22. The PXRD patterns for thesesalts indicate that the packing of the molecules within the crystallattice of these mixed salts were roughly equivalent. Minor changes inthe molecular packing density resulted to accommodate the differingratios of S and R camphor sulfonic acid in the lattice. This change inpacking density resulted in small shifts in peak position for certain ofthe peaks in the PXRD patterns. Camsylate salts containing differentratios of the R and S camphor sulfonic acid, to those described herein,could also be formed, and these salts would have roughly equivalentcrystal lattices.

The experimental PXRD pattern of the 1R:1S-camsylate salt, expressed interms of the degree (2θ) and relative intensities with a relativeintensity greater than 10.0%, measured on a Bruker-AXS Ltd., D4diffractometer with CuKα radiation at 1.5406 Å, is also shown in Table20.

TABLE 20 Angle Relative Intensity (Degree 2θ ± 0.1°) (>10.0%) 11.9 26.113.1 40.2 13.5 47.5 14.7 34.1 16.0 26.6 16.3 20.9 17.9 22.7 18.7 19.219.1 29.0 20.1 22.7 20.5 18.6 21.0 30.6 21.9 33.3 22.5 10.9 23.5 31.523.9 14.2 24.3 14.4 25.1 100.0 27.1 16.1 27.8 11.3 28.7 13.8 29.7 11.630.2 13.2 30.7 14.4

The 1R:1S-camsylate salt and the 1R:9S-camsylate salt were alsocharacterized by the solid state NMR spectral pattern shown in FIGS. 30and 32, carried out on a Bruker-Biospin 4 mm BL CPMAS probe positionedinto a wide-bore Bruker-Biospin DSX 500 MHz NMR spectrometer asdescribed in Example 24. The ¹³C chemical shifts of the 1R:1S-camsylatesalt is shown in Table 21.

TABLE 21 ¹³C Chemical Shifts^(a) [ppm] Intensity^(b) 214.3 4.8 212.2 0.6171.6 4.7 159.8 2.2 157.9 3.2 136.9 5.2 134.7 4.6 132.3 10.3 129.0 4.4128.0 8.3 126.6 4.5 124.8 4.7 123.2 4.6 112.9 5.9 110.7 3.5 102.7 2.758.7 7.8 52.5 4.6 49.6 0.9 48.0 12.0 42.4 8.8 40.5 0.9 36.1 5.8 31.5 4.228.4 3.7 24.7 4.4 20.5 6.3 18.2 7.6 ^(a)Referenced to external sample ofcrystalline adamantane at 29.5 ppm. ^(b)Defined as peak heights.Intensities can vary depending on the actual setup of the CPMASexperimental parameters and the thermal history of the sample. CPMASintensities are not necessarily quantitative.The ¹³C chemical shifts of the 1R:9S-camsylate salt is shown in Table22.

TABLE 22 ¹³C Chemical Shifts^(a) [ppm] Intensity^(b) 214.4 5.1 212.0 2.0171.8 6.0 159.9 3.1 158.0 4.2 137.1 6.8 134.8 4.5 132.5 12.0 128.3 10.8126.7 6.1 124.7 3.9 123.3 5.1 112.9 5.6 110.5 3.9 102.8 2.8 101.6 1.558.7 8.1 52.6 5.6 50.0 2.7 48.2 11.4 42.6 11.9 40.6^( c) 1.1 36.2 6.531.5 5.3 28.4 3.8 27.6^( c) 2.8 24.8 5.8 20.4 6.0 19.2^( c) 3.9 18.310.0 ^(a)Referenced to external sample of crystalline adamantane at 29.5ppm. ^(b)Defined as peak heights. Intensities can vary depending on theactual setup of the CPMAS experimental parameters and the thermalhistory of the sample. CPMAS intensities are not necessarilyquantitative. ^(c) Peak shoulder.

The 1R:1S-camsylate salt and the 1R:9S-camsylate salt were alsocharacterized by the solid state NMR spectral pattern shown in FIGS. 31and 33, carried out on a Bruker-Biospin 4 mm BL CPMAS probe positionedinto a wide-bore Bruker-Biospin DSX 500 MHz NMR spectrometer asdescribed in Example 23. The ¹⁹F chemical shifts of the 1R:1 S-camsylatesalt are shown in Table 23.

TABLE 23 ¹⁹F Chemical Shifts^(a) [ppm] Intensity^(b) −118.0 1.9 −119.912.0 −121.6 1.8 ^(a)Referenced to external standard of trifluoroaceticacid (50% V/V in H₂O) at −76.54 ppm. ^(b)Defined as peak heights.

The ¹⁹F chemical shifts of the 1R:9S-camsylate salt are shown in Table24.

TABLE 24 ¹⁹F Chemical Shifts^(a) [ppm] Intensity^(b) −117.9 2.0 −119.812.0 ^(a)Referenced to external sample of crystalline adamantane at 29.5ppm. bDefined as peak heights.

The DSC thermograms for the 1R:1S-camsylate salt and the 1R:9S-camsylatesalt, shown in FIGS. 25 and 26, indicate an endotherm onset at 303.2° C.for the 1R:1 S-camsylate salt and an endotherm onset at 301.2° C. forthe 1R:9S-camsylate salt.

The 1R:1S-camsylate salt was also characterized by FourierTransform-Infrared Spectroscopy (FT-IR) as described in Example 25, andthe spectral peaks are shown in Table 25. Absorption band frequenciesare listed. (w: weak, m: medium, s: strong, vs: very strong).Experimental error is ±2 cm⁻¹ except for * error on peak position couldbe considerably larger.

TABLE 25 Wavenumber (cm⁻¹) 3293 w 3078 w 2966 w 2915 w 1754 w 1743 s1635 m 1615 s 1582 w 1513 w 1475 m 1463 m 1446 s 1416 m 1366 w 1347 m1324 m 1315 m 1266 m 1254 m 1241 s 1216 m 1194 m 1180 m 1156 s 1132 s1125 s 1106 s 1066 m 1056 m 1028 s 982 w 964 w 959 w 950 w 937 w 899 w869 s 856 m 843 m 810 m 788 s 754 m 742 w 721 m 705 m 674 m 657 m

The 1R:1S-camsylate salt was also characterized by FourierTransform-Raman Spectroscopy (FT-Raman) as described in Example 26, andthe spectral peaks are shown in Table 26. (w: weak, m: medium, s:strong, vs: very strong). Experimental error is ±2 cm⁻¹.

TABLE 26 Wavenumber (cm⁻¹) 3298 w* 3228 w 3074 w 3059 w 3026 w 2986 w2965 w 2943 w 2917 w 2895 w 2845 w 2818 w 2777 w 2718 w 2553 w 1744 w1616 vs 1583 s 1578 s 1554 vs 1511 w 1454 vs 1434 m 1419 w 1407 w 1368 m1348 s 1324 m 1269 w 1251 w 1218 w 1204 m 1164 w 1135 w 1068 s 1039 w1021 w 1002 w 960 w 939 w 901 w 874 w 859 w 813 w 795 w 754 w 725 w 706w 678 w 646 w 616 w 581 w 549 w 515 w 504 w 485 w 443 w 430 w 413 w 400w 369 w 350 w 340 w 277 w 261 w 242 w 215 w 161 w 136 w 116 m 86 m 63 m

H. R-Camsylate Salt of Compound 1, R-Camsylate Polymorph Form A

The R-camsylate salt of Compound 1, R-camsylate polymorph Form A, can beproduced as described in Example 21.

R-camsylate polymorph Form A was characterized by measuring the PXRDpattern for a particular batch of R-camsylate polymorph Form A. Thisexperimental PXRD pattern is shown in FIG. 23. The experimental PXRDpattern of R-camsylate polymorph Form A, expressed in terms of thedegree (2θ) and relative intensities with a relative intensity greaterthan 10.0%, measured on a Bruker-AXS Ltd., D4 diffractometer with CuKαradiation at 1.5406 Å, is also shown in Table 27.

TABLE 27 Angle Relative Intensity (Degree 2θ ± 0.1°) (>10.0%) 6.1 33.212.2 76.9 12.7 60.6 13.5 67.0 13.8 16.0 14.3 29.9 14.8 39.8 15.2 12.315.9 22.9 16.1 24.1 16.6 29.7 18.2 49.3 18.5 34.9 19.1 24.4 19.5 37.220.5 58.9 21.0 56.3 22.4 52.9 23.0 24.6 23.4 14.8 24.0 47.7 24.5 25.925.0 21.6 25.6 100.0 26.5 13.5 27.4 17.5 28.4 16.6 29.0 14.0 29.8 14.430.7 18.5 31.5 10.1 32.9 14.3

The DSC thermogram for R-camsylate polymorph Form A, shown in FIG. 27,indicates an endotherm onset at 301.0° C.

R-camsylate polymorph Form A was also characterized by FourierTransform-Infrared Spectroscopy (FT-IR) as described in Example 25, andthe spectral peaks are shown in Table 28. Absorption band frequenciesare listed. (w: weak, m: medium, s: strong, vs: very strong).Experimental error is ±2 cm⁻¹ except for * error on peak position couldbe considerably larger.

TABLE 28 Wavenumber (cm⁻¹) 3288 m 3238 m 3076 w 2966 w 2949 w 2916 w2892 w 2840 w 1743 s 1637 s 1615 s 1581 w 1510 w 1474 m 1451 m 1416 m1366 w 1348 w 1315 m 1290 w 1266 m 1255 m 1240 s 1234 s 1203 s 1193 s1152 s 1129 s 1104 s 1066 m 1056 w 1029 s 979 w 967 w 958 w 951 w 937 w899 w 870 m 864 m 848 m 835 m 811 m 787 s 754 m 720 m 707 m 674 m 660 m

R-camsylate polymorph Form A was also characterized by FourierTransform-Raman Spectroscopy (FT-Raman) as described in Example 26, andthe spectral peaks are shown in Table 29. (w: weak, m: medium, s:strong, vs: very strong). Experimental error is ±2 cm⁻¹.

TABLE 29 Wavenumber (cm⁻¹) 3296 w 3231 w 3109 w 3075 w 3059 w 3042 w3024 w 2999 w 2966 w 2942 w 2921 w 2895 w 2845 w 2820 w 2777 w 2718 w2555 w 1745 w 1617 vs 1581 s 1554 vs 1510 w 1454 vs 1434 w 1419 w 1408 w1369 m 1348 m 1324 m 1270 w 1251 w 1214 w 1201 w 1160 w 1134 w 1068 s1041 w 1022 w 939 w 901 w 859 w 816 w 726 w 708 w 679 w 645 w 621 w 585w 549 w 516 w 503 w 484 w 430 w 415 w 370 w 350 w 277 w 261 w 243 w 219w 158 w 137 w 115 m 84 m 64 m

R-camsylate polymorph Forms B and C can also be produced andcharacterized according to the methods described above.

II. Amorphous Form of the S-Camsylate Salt of Compound 1

The amorphous form of the S-camsylate salt of Compound 1, can beproduced as described in Example 27.

The amorphous form of the S-camsylate salt of Compound 1 wascharacterized by measuring the PXRD pattern for a particular batch ofthe amorphous form of the S-camsylate salt of Compound 1, as describedin Example 28. This experimental PXRD pattern is shown in FIG. 34.

The amorphous form of the S-camsylate salt of Compound 1 was alsocharacterized by the solid state NMR spectral pattern shown in FIG. 35,carried out on a Bruker-Biospin 4 mm BL CPMAS probe positioned into awide-bore Bruker-Biospin DSX 500 MHz NMR spectrometer as described inExample 29. The ¹³C chemical shifts of the amorphous form of theS-camsylate salt of Compound 1 are shown in Table 30.

TABLE 30 ¹³C Chemical Shifts^(a) [ppm] Intensity^(b) 216.1 1.5 171.5 3.6160.2^( c) 2.8 158.6 3.4 137.7 6.6 131.8 9.2 129.5^( c) 7.3 124.3 6.2111.7 5.7 102.6 2.3 58.7 6.1 52.8 3.2 48.0 9.6 42.9 12.0 34.6 2.8 31.24.7 27.5 7.3 19.8 9.0 ^(a)Referenced to external sample of solid phaseadamantane at 29.5 ppm. ^(b)Defined as peak heights. Intensities canvary depending on the actual setup of the CPMAS experimental parametersand the thermal history of the sample. CPMAS intensities are notnecessarily quantitative. ^(c) Peak shoulder.

The amorphous form of the S-camsylate salt of Compound 1 was alsocharacterized by the solid state NMR spectral pattern shown in FIG. 36,carried out on a Bruker-Biospin 4 mm BL CPMAS probe positioned into awide-bore Bruker-Biospin DSX 500 MHz NMR spectrometer as described inExample 29. The ¹⁹F chemical shifts of the amorphous form of theS-camsylate salt of Compound 1 are shown in Table 31.

TABLE 31 ¹⁹F Chemical Shifts^(a) [ppm] Intensity^(b) −120.3 12.0^(a)Referenced to external standard of trifluoroacetic acid (50% V/V inH₂O) at −76.54 ppm. ^(b)Defined as peak heights.

The DSC thermogram for the amorphous form of the S-camsylate salt ofCompound 1, shown in FIG. 38, indicates a glass transition temperature(Tg) of 156.5° C.

III. Pharmaceutical Compositions of the Invention

The active agents (e.g., the crystalline salt forms or solid formscomprising two or more such forms, of Compound 1) described herein maybe formulated into pharmaceutical compositions suitable for mammalianmedical use. Any suitable route of administration may be employed forproviding a patient with an effective dosage of any of the polymorphicforms of Compound 1. For example, peroral or parenteral formulations andthe like may be employed. Dosage forms include capsules, tablets,dispersions, suspensions and the like, e.g. enteric-coated capsulesand/or tablets, capsules and/or tablets containing enteric-coatedpellets of Compound 1. In all dosage forms, polymorphic forms ofCompound 1 can be admixtured with other suitable constituents. Thecompositions may be conveniently presented in unit dosage forms, andprepared by any methods known in the pharmaceutical arts. Pharmaceuticalcompositions of the invention typically include a therapeuticallyeffective amount of the active agent and one or more inert,pharmaceutically acceptable carriers, and optionally any othertherapeutic ingredients, stabilizers, or the like. The carrier(s) aretypically pharmaceutically acceptable in the sense of being compatiblewith the other ingredients of the formulation and not unduly deleteriousto the recipient thereof. The compositions may further include diluents,buffers, binders, disintegrants, thickeners, lubricants, preservatives(including antioxidants), flavoring agents, taste-masking agents,inorganic salts (e.g., sodium chloride), antimicrobial agents (e.g.,benzalkonium chloride), sweeteners, antistatic agents, surfactants(e.g., polysorbates such as “TWEEN 20” and “TWEEN 80”, and pluronicssuch as F68 and F88, available from BASF), sorbitan esters, lipids(e.g., phospholipids such as lecithin and other phosphatidylcholines,phosphatidylethanolamines, fatty acids and fatty esters, steroids (e.g.,cholesterol)), and chelating agents (e.g., EDTA, zinc and other suchsuitable cations). Other pharmaceutical excipients and/or additivessuitable for use in the compositions according to the invention arelisted in Remington: The Science & Practice of Pharmacy, 19^(th) ed.,Williams & Williams, (1995), and in the “Physician's Desk Reference”,52^(nd) ed., Medical Economics, Montvale, N.J. (1998), and in Handbookof Pharmaceutical Excipients, 3^(rd). Ed., Ed. A. H. Kibbe,Pharmaceutical Press, 2000. The active agents of the invention may beformulated in compositions including those suitable for oral, rectal,topical, nasal, ophthalmic, or parenteral (including intraperitoneal,intravenous, subcutaneous, or intramuscular injection) administration.

The amount of the active agent in the formulation can vary dependingupon a variety of factors, including dosage form, the condition to betreated, target patient population, and other considerations, and willgenerally be readily determined by one skilled in the art. Atherapeutically effective amount will typically be an amount necessaryto modulate, regulate, or inhibit a PARP enzyme. In practice, this canvary widely depending, for example, upon the particular active agent,the severity of the condition to be treated, the patient population, thestability of the formulation, and the like. Compositions will generallycontain anywhere from about 0.001% by weight to about 99% by weightactive agent, preferably from about 0.01% to about 5% by weight activeagent, and more preferably from about 0.01% to 2% by weight activeagent, and can also depend upon the relative amounts ofexcipients/additives contained in the composition.

In some embodiments, a pharmaceutical composition can be administered inconventional dosage form prepared by combining a therapeuticallyeffective amount of an active agent as an active ingredient with one ormore appropriate pharmaceutical carriers according to conventionalprocedures. These procedures may involve mixing, granulating andcompressing or dissolving the ingredients as appropriate to the desiredpreparation.

The pharmaceutical carrier(s) employed may be either solid or liquid.Exemplary solid carriers include, but are not limited to, lactose,sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate,stearic acid and the like. Exemplary liquid carriers include syrup,peanut oil, olive oil, water and the like. Similarly, the carrier(s) mayinclude time-delay or time-release materials known in the art, such asglyceryl monostearate or glyceryl distearate alone or with a wax,ethylcellulose, hydroxypropylmethylcellulose, methylmethacrylate and thelike.

A variety of pharmaceutical forms can be employed. For example, if asolid carrier is used, the preparation can be tableted, placed in a hardgelatin capsule in powder or pellet form or in the form of a troche orlozenge. The amount of solid carrier may vary, but generally will befrom about 25 mg to about 1 g. If a liquid carrier is used, thepreparation can be in the form of syrup, emulsion, soft gelatin capsule,sterile injectable solution or suspension in an ampoule or vial ornon-aqueous liquid suspension.

To obtain a stable water-soluble dose form, a pharmaceuticallyacceptable salt of an active agent can be dissolved in an aqueoussolution of an organic or inorganic acid, such as 0.3 M solution ofsuccinic acid or citric acid. If a soluble salt form is not available,the active agent may be dissolved in a suitable co-solvent orcombinations of co-solvents. Examples of suitable co-solvents include,but are not limited to, alcohol, propylene glycol, polyethylene glycol300, polysorbate 80, gylcerin and the like in concentrations rangingfrom about 0 to about 60% of the total volume. The composition may alsobe in the form of a solution of a salt form of the active agent in anappropriate aqueous vehicle such as water or isotonic saline or dextrosesolution.

It will be appreciated that the actual dosages of Compound 1 used in thecompositions of this invention can vary according to the particularpolymorphic form being used, the particular composition formulated, themode of administration and the particular site, host and disease beingtreated. Those skilled in the art using conventionaldosage-determination tests in view of the experimental data for an agentcan ascertain optimal dosages for a given set of conditions. For oraladministration, an exemplary daily dose generally employed is from about0.001 to about 1000 mg/kg of body weight, more preferably from about0.001 to about 50 mg/kg body weight, and courses of treatment can berepeated at appropriate intervals. Administration of prodrugs istypically dosed at weight levels that are chemically equivalent to theweight levels of the fully active form. In the practice of theinvention, the most suitable route of administration as well as themagnitude of a therapeutic dose will depend on the nature and severityof the disease to be treated. The dose, and dose frequency, may alsovary according to the age, body weight, and response of the individualpatient. In general, a suitable oral dosage form may cover a dose rangefrom 0.5 mg to 100 mg of active ingredient total daily dose,administered in one single dose or equally divided doses. A preferredamount of Compound 1 in such formulations is from about 0.5 mg to about20 mg, such as from about 1 mg to about 10 mg or from about 1 mg toabout 5 mg.

The compositions of the invention may be manufactured in mannersgenerally known for preparing pharmaceutical compositions, e.g., usingconventional techniques such as mixing, dissolving, granulating,dragee-making, levigating, emulsifying, encapsulating, entrapping orlyophilizing. Pharmaceutical compositions may be formulated in aconventional manner using one or more physiologically acceptablecarriers, which may be selected from excipients and auxiliaries thatfacilitate processing of the active compounds into preparations that canbe used pharmaceutically.

For oral administration, a polymorphic form of Compound 1 can beformulated readily by combining the active agent with pharmaceuticallyacceptable carriers known in the art. Such carriers enable the compoundsof the invention to be formulated as tablets, pills, dragees, capsules,gels, syrups, slurries, suspensions and the like, for oral ingestion bya patient to be treated. Pharmaceutical preparations for oral use can beobtained using a solid excipient in admixture with the active agent,optionally grinding the resulting mixture, and processing the mixture ofgranules after adding suitable auxiliaries, if desired, to obtaintablets or dragee cores. Suitable excipients include: fillers such assugars, including lactose, sucrose, mannitol, or sorbitol; and cellulosepreparations, for example, maize starch, wheat starch, rice starch,potato starch, gelatin, gum, methyl cellulose,hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, orpolyvinylpyrrolidone (PVP). If desired, disintegrating agents may beadded, such as crosslinked polyvinyl pyrrolidone, agar, or alginic acidor a salt thereof such as sodium alginate.

Dragee cores are provided with suitable coatings. For this purpose,concentrated sugar solutions may be used, which may optionally containgum arabic, polyvinyl pyrrolidone, Carbopol gel, polyethylene glycol,and/or titanium dioxide, lacquer solutions, and suitable organicsolvents or solvent mixtures. Dyestuffs or pigments may be added to thetablets or dragee coatings for identification or to characterizedifferent combinations of active agents.

Pharmaceutical preparations that can be used orally include push-fitcapsules made of gelatin, as well as soft, sealed capsules made ofgelatin and a plasticizer, such as glycerol or sorbitol. The push-fitcapsules can contain the active ingredients in admixture with fillerssuch as lactose, binders such as starches, and/or lubricants such astalc or magnesium stearate, and, optionally, stabilizers. In softcapsules, the active agents may be dissolved or suspended in suitableliquids, such as fatty oils, liquid paraffin, or liquid polyethyleneglycols. In addition, stabilizers may be added. All formulations fororal administration should be in dosages suitable for suchadministration. For buccal administration, the compositions may take theform of tablets or lozenges formulated in conventional manner.

For administration intranasally or by inhalation, the compounds can beconveniently delivered in the form of an aerosol spray presentation frompressurized packs or a nebuliser, with the use of a suitable propellant,e.g., dichlorodifluoromethane, trichlorofluoromethane,dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In thecase of a pressurized aerosol the dosage unit may be determined byproviding a valve to deliver a metered amount. Capsules and cartridgesof gelatin for use in an inhaler or insufflator and the like may beformulated containing a powder mix of the compound and a suitable powderbase such as lactose or starch.

The active agents may be formulated for parenteral administration byinjection, e.g., by bolus injection or continuous infusion. Formulationsfor injection may be presented in unit-dosage form, e.g., in ampoules orin multi-dose containers, with an added preservative. The compositionsmay take such forms as suspensions, solutions or emulsions in oily oraqueous vehicles, and may contain formulatory agents such as suspending,stabilizing and/or dispersing agents.

Pharmaceutical formulations for parenteral administration include, forexample, suspensions of the active agents and may be prepared asappropriate oily injection suspensions. Suitable lipophilic solvents orvehicles include fatty oils such as sesame oil, or synthetic fatty acidesters, such as ethyl oleate or triglycerides, or liposomes. Aqueousinjection suspensions may contain substances that increase the viscosityof the suspension, such as sodium carboxymethyl cellulose, sorbitol, ordextran. Optionally, the suspension may also contain suitablestabilizers or agents that increase the solubility of the active agentsto allow for the preparation of highly concentrated solutions.

For administration to the eye, the active agent can be delivered in apharmaceutically acceptable ophthalmic vehicle such that the compound ismaintained in contact with the ocular surface for a sufficient timeperiod to allow the compound to penetrate the corneal and internalregions of the eye, including, for example, the anterior chamber,posterior chamber, vitreous body, aqueous humor, vitreous humor, cornea,iris/cilary, lens, choroid/retina and selera. The pharmaceuticallyacceptable ophthalmic vehicle may be, for example, an ointment,vegetable oil, or an encapsulating material. An active agent of theinvention may also be injected directly into the vitreous and aqueoushumor or subtenon.

Alternatively, the active ingredient may be in powder form forconstitution with a suitable vehicle, e.g., sterile pyrogen-free water,before use. The compounds may also be formulated in rectal or vaginalcompositions such as suppositories or retention enemas, e.g., containingconventional suppository bases such as cocoa butter or other glycerides.

In addition to the formulations described above, the polymorphic formsmay also be formulated as a depot preparation. Such long-actingformulations may be administered by implantation (for example,subcutaneously or intramuscularly) or by intramuscular injection. Thus,for example, the polymorphic forms may be formulated with suitablepolymeric or hydrophobic materials (for example, as an emulsion in anacceptable oil) or ion-exchange resins, or as sparingly solublederivatives, for example, as a sparingly soluble salt.

Additionally, polymorphic forms of Compound 1 may be delivered using asustained-release system, such as semipermeable matrices of solidhydrophobic polymers containing the therapeutic agent. Varioussustained-release materials have been established and are known by thoseskilled in the art. Sustained-release capsules may, depending on theirchemical nature, release the compound for a few weeks up to over 100days.

The pharmaceutical compositions also may comprise suitable solid- orgel-phase carriers or excipients. Examples of such carriers orexcipients include calcium carbonate, calcium phosphate, sugars,starches, cellulose derivatives, gelatin, and polymers such aspolyethylene glycols.

IV. Methods of Using the Polymorphs of the Invention

Polymorphic forms of the crystalline salts of Compound 1 can be usefulfor mediating the activity of poly(ADP-ribose) polymerase (PARP). Moreparticularly, these polymorphic forms can be useful as chemosensitizersthat enhance the efficacy of radiotherapy or cytotoxic drugs whosemechanism depends on DNA damage. These drugs include, but are notlimited to, temozolomide (SCHERING), irinotecan (PFIZER), topotecan(GLAXO SMITHKLINE), cisplatin (BRISTOL MEYERS SQUIBB; AM PHARM PARTNERS;BEDFORD; GENSIA SICOR PHARMS; PHARMACHEMIE), and doxorubicinhydrochloride (AM PHARM PARTNERS; BEDFORD; GENSIA; SICOR PHARMS;PHARMACHEMIE; ADRIA; ALZA).

Polymorphic salt forms of Compound 1 can also be useful for enhancingthe induction of the expression of Reg gene in 1 cells and HGF gene and,accordingly, promoting the proliferation of pancreatic 1-cells ofLangerhans' islets and suppressing apoptosis of the cells. Further, theinventive polymorphic salt forms of Compound 1 can be useful forpreparing cosmetics, for example, in after-sun lotions.

Therapeutically effective amounts of the agents of the invention may beadministered, typically in the form of a pharmaceutical composition, totreat diseases mediated by modulation or regulation of PARP. An“effective amount” refers to that amount of an agent that, whenadministered to a mammal, including a human, in need of such treatment,is sufficient to effect treatment for a disease mediated by the activityof one or more PARP enzyme. Thus, a therapeutically effective amount ofa compound refers to a quantity sufficient to modulate, regulate, orinhibit the activity of one or more PARP enzyme such that a diseasecondition that is mediated by that activity is reduced or alleviated.The effective amount of a given compound can vary depending upon factorssuch as the disease condition and its severity and the identity andcondition (e.g., weight) of the mammal in need of treatment, but cannevertheless be routinely determined by one skilled in the art.“Treating” refers to at least the mitigation of a disease condition in amammal, including a human, that is affected, at least in part, by theactivity of one or more PARP enzymes and includes: preventing thedisease condition from occurring in a mammal, particularly when themammal is found to be predisposed to having the disease condition buthas not yet been diagnosed as having it; modulating and/or inhibitingthe disease condition; and/or alleviating the disease condition.Exemplary disease conditions include diabetic retinopathy, neovascularglaucoma, rheumatoid arthritis, psoriasis, age-related maculardegeneration (AMD), and abnormal cell growth, such as cancer. Cancerincludes, but is not limited to, mesothelioma, hepatobilliary (hepaticand billiary duct), a primary or secondary CNS tumor, a primary orsecondary brain tumor, lung cancer (NSCLC and SCLC), bone cancer,pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous orintraocular melanoma, ovarian cancer, colon cancer, rectal cancer,cancer of the anal region, stomach cancer, gastrointestinal (gastric,colorectal, and duodenal), breast cancer, uterine cancer, carcinoma ofthe fallopian tubes, carcinoma of the endometrium, carcinoma of thecervix, carcinoma of the vagina, carcinoma of the vulva, Hodgkin'sDisease, cancer of the esophagus, cancer of the small intestine, cancerof the endocrine system, cancer of the thyroid gland, cancer of theparathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue,cancer of the urethra, cancer of the penis, prostate cancer, testicularcancer, chronic or acute leukemia, chronic myeloid leukemia, lymphocyticlymphomas, cancer of the bladder, cancer of the kidney or ureter, renalcell carcinoma, carcinoma of the renal pelvis, neoplasms of the centralnervous system (CNS), primary CNS lymphoma, non hodgkins's lymphoma,spinal axis tumors, brain stem glioma, pituitary adenoma, adrenocorticalcancer, gall bladder cancer, multiple myeloma, cholangiocarcinoma,fibrosarcoma, neuroblastoma, retinoblastoma, or a combination of one ormore of the foregoing cancers.

Abnormal cell growth also includes, but is not limited to, benignproliferative diseases, including, but not limited to, psoriasis, benignprostatic hypertrophy or restinosis.

The activity of the polymorphic salt forms of Compound 1 as modulatorsof PARP activity may be measured by any of the methods available tothose skilled in the art, including in vivo and/or in vitro assays.Examples of suitable assays for activity measurements include thosedescribed in U.S. Pat. No. 6,495,541 and U.S. Provisional PatentApplication No. 60/612,458, the disclosures of which are incorporatedherein by reference in their entireties.

Some embodiments are also directed to therapeutic methods of treating adisease condition mediated by PARP activity, for example, cancer and avariety of disease and toxic states that involve oxidative or nitricoxide induced stress and subsequent PARP hyperactivation. Suchconditions include, but are not limited to, neurologic andneurodegenerative disorders (eg, Parkinson's disease, Alzheimer'sdisease), cardiovascular disorders (e.g., myocardial infarction,ischemia-reperfusion injury), diabetic vascular dysfunction,cisplatin-induced nephrotoxicity. In some embodiments, the therapeuticmethods include administering to a mammal in need thereof atherapeutically effective amount of a pharmaceutical composition whichincludes any of the polymorphic forms, or pharmaceutical compositionsdiscussed herein.

Some embodiments are also directed to combination therapeutic methods oftreating a disease condition mediated by PARP activity, which comprisesadministering to a mammal in need thereof a therapeutically effectiveamount of a pharmaceutical composition which comprises any of thepolymorphic forms, or pharmaceutical compositions discussed herein, incombination with a therapeutically effective amount of one or moresubstances, such as anti-tumor agents, anti-angiogenesis agents, signaltransduction inhibitors, and antiproliferative agents, mitoticinhibitors, alkylating agents, anti-metabolites, intercalatingantibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes,topoisomerase inhibitors, biological response modifiers, antibodies,cytotoxics, anti-hormones, and anti-androgens. Such substances include,but are not limited to, those disclosed in PCT Publication Nos. WO00/38715, WO 00/38716, WO 00/38717, WO 00/38718, WO 00/38719, WO00/38730, WO 00/38665, WO 00/37107 and WO 00/38786, the disclosures ofwhich are incorporated herein by reference in their entireties.

Examples of anti-tumor agents include temozolomide (SCHERING),irinotecan (PFIZER), topotecan (GLAXO SMITHKLINE), cisplatin (BRISTOLMEYERS SQUIBB; AM PHARM PARTNERS; BEDFORD; GENSIA SICOR PHARMS;PHARMACHEMIE), and doxorubicin hydrochloride (AM PHARM PARTNERS;BEDFORD; GENSIA; SICOR PHARMS; PHARMACHEMIE; ADRIA; ALZA).

Additional examples of anti-tumor agents include mitotic inhibitors, forexample vinca alkaloid derivatives such as vinblastine vinorelbine,vindescine and vincristine; colchines allochochine, halichondrine,N-benzoyltrimethyl-methyl ether colchicinic acid, dolastatin 10,maystansine, rhizoxine, taxanes such as taxol (paclitaxel), docetaxel(Taxotere), 2′-N-[3-(dimethylamino)propyl]glutaramate (taxolderivative), thiocholchicine, trityl cysteine, teniposide, methotrexate,azathioprine, fluorouricil, cytocine arabinoside,2′2′-difluorodeoxycytidine (gemcitabine), adriamycin and mitamycin.Alkylating agents, for example, carboplatin, oxiplatin, iproplatin,ethyl ester of N-acetyl-DL-sarcosyl-L-leucine (Asaley or Asalex),1,4-cyclohexadiene-1,4-dicarbamic acid, 2,5-bis(1-azirdinyl)-3,6-dioxo-,diethyl ester (diaziquone), 1,4-bis(methanesulfonyloxy)butane (bisulfanor leucosulfan), chlorozotocin, clomesone, cyanomorpholinodoxorubicin,cyclodisone, dianhydroglactitol, fluorodopan, hepsulfam, mitomycin C,hycantheonemitomycin C, mitozolamide,1-(2-chloroethyl)-4-(3-chloropropyl)-piperazine dihydrochloride,piperazinedione, pipobroman, porfiromycin, spirohydantoin mustard,teroxirone, tetraplatin, thiotepa, triethylenemelamine, uracil nitrogenmustard, bis(3-mesyloxypropyl)amine hydrochloride, mitomycin,nitrosoureas agents such as cyclohexyl-chloroethylnitrosourea,methylcyclohexyl-chloroethylnitrosourea,1-(2-chloroethyl)-3-(2,6-dioxo-3-piperidyl)-1-nitroso-urea,bis(2-chloroethyl)nitrosourea, procarbazine, dacarbazine, nitrogenmustard-related compounds such as mechloroethamine, cyclophosphamide,ifosamide, melphalan, chlorambucil, estramustine sodium phosphate, andstrptozoin. DNA anti-metabolites, for example 5-fluorouracil, cytosinearabinoside, hydroxyurea,2-[(3hydroxy-2-pyrinodinyl)methylene]-hydrazinecarbothioamide,deoxyfluorouridine, 5-hydroxy-2-formylpyridine thiosemicarbazone,alpha-2′-deoxy-6-thioguanosine, aphidicolin glycinate,5-azadeoxycytidine, beta-thioguanine deoxyriboside, cyclocytidine,guanazole, inosine glycodialdehyde, macbecin II, pyrazolimidazole,cladribine, pentostatin, thioguanine, mercaptopurine, bleomycin,2-chlorodeoxyadenosine, inhibitors of thymidylate synthase such asraltitrexed and pemetrexed disodium, clofarabine, floxuridine andfludarabine. DNA/RNA antimetabolites, for example, L-alanosine,5-azacytidine, acivicin, aminopterin and derivatives thereof such asN-[2-chloro-5-[[(2,4-diamino-5-methyl-6-quinazolinyl)methyl]amino]benzoyl]-L-aspartic acid,N-[4-[[(2,4-diamino-5-ethyl-6-quinazolinyl)methyl]amino]benzoyl]-L-aspartic acid,N-[2-chloro-4-[[(2, 4-diaminopteridinyl)methyl]amino]benzoyl]-L-asparticacid, soluble Baker's antifol, dichloroallyl lawsone, brequinar, ftoraf,dihydro-5-azacytidine, methotrexate, N-(phosphonoacetyl)-L-aspartic acidtetrasodium salt, pyrazofuran, trimetrexate, plicamycin, actinomycin D,cryptophycin, and analogs such as cryptophycin-52 or, for example, oneof the preferred anti-metabolites disclosed in European PatentApplication No. 239362 such asN-(5-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-methylamino]-2-thenoyl)-L-glutamicacid; growth factor inhibitors; cell cycle inhibitors; intercalatingantibiotics, for example adriamycin and bleomycin; proteins, for exampleinterferon; and anti-hormones, for example anti-estrogens such asNolvadex™ (tamoxifen) or, for example anti-androgens such as Casodex™(4′-cyano-3-(4-fluorophenylsulphonyl)-2-hydroxy-2-methyl-3′-(trifluoromethyl)propionanilide).Such conjoint treatment may be achieved by way of the simultaneous,sequential or separate dosing of the individual components of thetreatment.

Anti-angiogenesis agents include MMP-2 (matrix-metalloprotienase 2)inhibitors, MMP-9 (matrix-metalloprotienase 9) inhibitors, and COX-II(cyclooxygenase II) inhibitors. Examples of useful COX-II inhibitorsinclude CELEBREX™ (alecoxib), valdecoxib, and rofecoxib. Examples ofuseful matrix metalloproteinase inhibitors are described in WO 96/33172(published Oct. 24, 1996), WO 96/27583 (published Mar. 7, 1996),European Patent Application No. 97304971.1 (filed Jul. 8, 1997),European Patent Application No. 99308617.2 (filed Oct. 29, 1999), WO98/07697 (published Feb. 26, 1998), WO 98/03516 (published Jan. 29,1998), WO 98/34918 (published Aug. 13, 1998), WO 98/34915 (publishedAug. 13, 1998), WO 98/33768 (published Aug. 6, 1998), WO 98/30566(published Jul. 16, 1998), European Patent Publication 606,046(published Jul. 13, 1994), European Patent Publication 931,788(published Jul. 28, 1999), WO 90/05719 (published May 331, 1990), WO99/52910 (published Oct. 21, 1999), WO 99/52889 (published Oct. 21,1999), WO 99/29667 (published Jun. 17, 1999), PCT InternationalApplication No. PCT/IB98/01113 (filed Jul. 21, 1998), European PatentApplication No. 99302232.1 (filed Mar. 25, 1999), Great Britain patentapplication number 9912961.1 (filed Jun. 3, 1999), U.S. ProvisionalApplication No. 60/148,464 (filed Aug. 12, 1999), U.S. Pat. No.5,863,949 (issued Jan. 26, 1999), U.S. Pat. No. 5,861,510 (issued Jan.19, 1999), and European Patent Publication 780,386 (published Jun. 25,1997), all of which are herein incorporated by reference in theirentirety. Preferred MMP-2 and MMP-9 inhibitors are those that havelittle or no activity inhibiting MMP-1. More preferred, are those thatselectively inhibit MMP-2 and/or MMP-9 relative to the othermatrix-metalloproteinases (i.e. MMP-1, MMP-3, MMP-4, MMP-5, MMP-6,MMP-7, MMP-8, MMP-10, MMP-11, MMP-12, and MMP-13).

Examples of MMP inhibitors include AG-3340, RO 32-3555, RS 13-0830, andthe following compounds:3-[[4-(4-fluoro-phenoxy)-benzenesulfonyl]-(1-hydroxycarbamoyl-cyclopentyl)-amino]-propionicacid;3-exo-3-[4-(4-fluoro-phenoxy)-benzenesulfonylamino]-8-oxa-bicyclo[3.2.1]octane-3-carboxylicacid hydroxyamide; (2R, 3R)1-[4-(2-chloro-4-fluoro-benzyloxy)-benzenesulfonyl]-3-hydroxy-3-methyl-piperidine-2-carboxylicacid hydroxyamide;4-[4-(4-fluoro-phenoxy)-benzenesulfonylamino]-tetrahydro-pyran-4-carboxylicacid hydroxyamide;3-[[4-(4-fluoro-phenoxy)-benzenesulfonyl]-(1-hydroxycarbamoyl-cyclobutyl)-amino]-propionicacid;4-[4-(4-chloro-phenoxy)-benzenesulfonylamino]-tetrahydro-pyran-4-carboxylicacid hydroxyamide;3-[4-(4-chloro-phenoxy)-benzenesulfonylamino]-tetrahydro-pyran-3-carboxylicacid hydroxyamide; (2R, 3R)1-[4-(4-fluoro-2-methyl-benzyloxy)-benzenesulfonyl]-3-hydroxy-3-methyl-piperidine-2-carboxylicacid hydroxyamide;3-[[4-(4-fluoro-phenoxy)-benzenesulfonyl]-(1-hydroxycarbamoyl-1-methyl-ethyl)-amino]-propionicacid;3-[[4-(4-fluoro-phenoxy)-benzenesulfonyl]-(4-hydroxycarbamoyl-tetrahydro-pyran-4-yl)-amino]-propionicacid;3-exo-3-[4-(4-chloro-phenoxy)-benzenesulfonylamino]-8-oxa-bicyclo[3.2.1]octane-3-carboxylicacid hydroxyamide;3-endo-3-[4-(4-fluoro-phenoxy)-benzenesulfonylamino]-8-oxa-bicyclo[3.2.1]octane-3-carboxylicacid hydroxyamide;3-[4-(4-fluoro-phenoxy)-benzenesulfonylamino]-tetrahydro-furan-3-carboxylicacid hydroxyamide; and pharmaceutically acceptable salts, solvates andhydrates thereof.

Examples of signal transduction inhibitors include agents that caninhibit EGFR (epidermal growth factor receptor) responses, such as EGFRantibodies, EGF antibodies, and molecules that are EGFR inhibitors; VEGF(vascular endothelial growth factor) inhibitors; and erbB2 receptorinhibitors, such as organic molecules or antibodies that bind to theerbB2 receptor, for example, HERCEPTIN™ (Genentech, Inc. of South SanFrancisco, Calif., USA).

EGFR inhibitors include, for example, those described in WO 95/19970(published Jul. 27, 1995), WO 98/14451 (published Apr. 9, 1998), WO98/02434 (published Jan. 22, 1998), and U.S. Pat. No. 5,747,498 (issuedMay 5, 1998). EGFR-inhibiting agents include, but are not limited to,the monoclonal antibodies C225 and anti-EGFR 22Mab (ImClone SystemsIncorporated of New York, N.Y., USA), the compounds ZD-1839(AstraZeneca), BIBX-1382 (Boehringer Ingelheim), MDX-447 (Medarex Inc.of Annandale, N.J., USA), and OLX-103 (Merck & Co. of WhitehouseStation, N.J., USA), VRCTC-310 (Ventech Research) and EGF fusion toxin(Seragen Inc. of Hopkinton, Mass.).

VEGF inhibitors, for example SU-5416 and SU-6668 (Sugen Inc. of SouthSan Francisco, Calif., USA), can also be combined or co-administeredwith the composition. Examples of VEGF inhibitors are described in, forexample in WO 99/24440 (published May 20, 1999), PCT InternationalApplication PCT/IB99/00797 (filed May 3, 1999), in WO 95/21613(published Aug. 17, 1995), WO 99/61422 (published Dec. 2, 1999), U.S.Pat. No. 5,834,504 (issued Nov. 10, 1998), WO 98/50356 (published Nov.12, 1998), U.S. Pat. No. 5,883,113 (issued Mar. 16, 1999), U.S. Pat. No.5,886,020 (issued Mar. 23, 1999), U.S. Pat. No. 5,792,783 (issued Aug.11, 1998), WO 99/10349 (published Mar. 4, 1999), WO 97/32856 (publishedSep. 12, 1997), WO 97/22596 (published Jun. 26, 1997), WO 98/54093(published Dec. 3, 1998), WO 98/02438 (published Jan. 22, 1998), WO99/16755 (published Apr. 8, 1999), and WO 98/02437 (published Jan. 22,1998), all of which are herein incorporated by reference in theirentirety. Other examples of some specific VEGF inhibitors are IM862(Cytran Inc. of Kirkland, Wash., USA); anti-VEGF monoclonal antibodybevacizumab (Genentech, Inc. of South San Francisco, Calif.); andangiozyme, a synthetic ribozyme from Ribozyme (Boulder, Colo.) andChiron (Emeryville, Calif.).

ErbB2 receptor inhibitors, such as GW-282974 (Glaxo Wellcome plc), andthe monoclonal antibodies AR-209 (Aronex Pharmaceuticals Inc. of TheWoodlands, Tex., USA) and 2B-1 (Chiron), may be administered incombination with the composition. Such erbB2 inhibitors include, but arenot limited to, those described in WO 98/02434 (published Jan. 22,1998), WO 99/35146 (published Jul. 15, 1999), WO 99/35132 (publishedJul. 15, 1999), WO 98/02437 (published Jan. 22, 1998), WO 97/13760(published Apr. 17, 1997), WO 95/19970 (published Jul. 27, 1995), U.S.Pat. No. 5,587,458 (issued Dec. 24, 1996), and U.S. Pat. No. 5,877,305(issued Mar. 2, 1999), each of which is herein incorporated by referencein its entirety. ErbB2 receptor inhibitors useful in the presentinvention are also described in U.S. Provisional Application No.60/117,341, filed Jan. 27, 1999, and in U.S. Provisional Application No.60/117,346, filed Jan. 27, 1999, both of which are herein incorporatedby reference in their entirety.

Other antiproliferative agents that may be used include, but are notlimited to, inhibitors of the enzyme farnesyl protein transferase andinhibitors of the receptor tyrosine kinase PDGFr, including thecompounds disclosed and claimed in the following U.S. patent applicationSer. No. 09/221,946 (filed Dec. 28, 1998); Ser. No. 09/454,058 (filedDec. 2, 1999); Ser. No. 09/501,163 (filed Feb. 9, 2000); Ser. No.09/539,930 (filed Mar. 31, 2000); Ser. No. 09/202,796 (filed May 22,1997); Ser. No. 09/384,339 (filed Aug. 26, 1999); and Ser. No.09/383,755 (filed Aug. 26, 1999); and the compounds disclosed andclaimed in the following United States provisional patent applications:60/168,207 (filed Nov. 30, 1999); 60/170,119 (filed Dec. 10, 1999);60/177,718 (filed Jan. 21, 2000); 60/168,217 (filed Nov. 30, 1999), and60/200,834 (filed May 1, 2000). Each of the foregoing patentapplications and provisional patent applications is herein incorporatedby reference in their entirety.

Compositions of the invention can also be used with other agents usefulin treating abnormal cell growth or cancer, including, but not limitedto, agents capable of enhancing antitumor immune responses, such asCTLA4 (cytotoxic lymphocite antigen 4) antibodies, and other agentscapable of blocking CTLA4; and anti-proliferative agents such as otherfarnesyl protein transferase inhibitors. Specific CTLA4 antibodies thatcan be used in the present invention include those described in U.S.Provisional Application 60/113,647 (filed Dec. 23, 1998), which isherein incorporated by reference in its entirety.

The disclosures of all cited references are incorporated herein byreference in their entirety.

EXAMPLES

The examples which follow will further illustrate the preparation andcharacterization of the distinct polymorphic salt forms of Compound 1,but are not intended to limit the scope of the invention as describedherein or as claimed herein. Unless otherwise indicated, alltemperatures are set forth in degrees Celsius and all parts andpercentages are by weight.

Example 1: Preparation of a Maleate Salt of Compound 1, MaleatePolymorph Form A

A solution of Compound 1 (100.8 mg; 0.31 mmol) in 80/20 v/v isopropylalcohol/water (25 mL) was prepared by dissolving the solid in the liquidmedium with stirring at ambient conditions (20-25° C.). A solution ofmaleic acid (25.13 mg; 0.22 mmol) in a minimum volume of 80/20 v/visopropyl alcohol/water was prepared as above. 17.26 mL of the solutionof Compound 1 was added slowly to the maleic acid solution with stirringat ambient conditions to provide an equimolar solution of Compound 1 andmaleic acid. The resulting solution was allowed to stir for 24 hours atambient conditions, followed by the addition of hexane (6 mL) andstorage at −20° C. for 24 hours; crystallization occurred during thattime. Following filtration and washing with 80/20 v/v isopropylalcohol/water, the product was dried under vacuum at 40° C. whichprovided approximately 100 mg of crystalline material.

Example 2: Preparation of a Maleate Salt of Compound 1, MaleatePolymorph Form B, Using Ethanol

A solution of Compound 1 (10 g; 30.9 mmol) in ethanol (450 mL) wasprepared by heating to reflux in a jacketed reaction vessel withoverhead stirring. A solution of maleic acid (3.95 g, 1.1 eq) in ethanol(20 mL) was added dropwise over 1 hour at 80° C.; crystallizationoccurred during this time. The suspension was cooled at 0.5° C./min andisolated at 0° C. after 1 hour granulation. Following filtration andwashing with ethanol (50 mL), the product was dried under vacuum at 50°C. to furnish 12 g of the crystalline product (89% theoretical yield).

Example 3: Preparation of a Maleate Salt of Compound 1, MaleatePolymorph Form B, Using Isopropyl Alcohol

A solution of Compound 1 (18 g; 55.7 mmol) in isopropyl alcohol (1500mL) was prepared by heating in a jacketed reaction vessel with overheadstirring. A solution of maleic acid (7.11 g, 1.1 eq) in isopropylalcohol (100 mL) was prepared and was added dropwise (over 1 hour)following the addition of seed crystals of the title compound (45 mg).Once the addition was complete, the suspension was cooled to 0° C. (atnatural rate) and granulated for 2 days. Following filtration theproduct was dried under vacuum at 50° C. to furnish 23.7 g of acrystalline product (97% theoretical yield).

Example 4: Preparation of an S-Camsylate Salt of Compound 1, S-CamsylatePolymorph Form A, Using Tetrahydrofuran

Compound 1 (20 g) was slurried at reflux in tetrahydrofuran (42 mL) andwater (40 mL) in a jacketed reaction vessel with overhead stirring, andremained as a free base slurry. S-camphor sulfonic acid solution (17.25g in 20 mL of water) was added slowly over approximately 10 minutes, toform a clear yellow solution, which was held at reflux for 30 minutes.Water (135 mL) was then added, over approximately 20 minutes,maintaining reflux. The resulting yellowish slurry was cooled to 10° C.and granulated at this temperature to improve crystallinity and yieldfor a suitable amount of time. Suitable granulation times can be chosenby one of skill in the art. Typical granulation times can range, forexample, from about 1 hour to about 48 hours. Filtered solids werewashed with chilled water (20 mL) and dried under vacuum at 50° C. togive the final product.

Example 5: Preparation of an S-Camsylate Salt of Compound 1, S-CamsylatePolymorph Form A, Using Isopropyl Alcohol

A solution of Compound 1 (982.5 mg; 3.03 mmol) in isopropyl alcohol (225mL) was prepared by dissolving the solid in the liquid medium withstirring at ambient conditions (20-25° C.). A solution of S-camphorsulfonic acid (53.81 mg) in a minimum volume of isopropyl alcohol wasprepared as above. 17.16 mL of the Compound 1 solution was added slowlyto the maleic acid solution with stirring at ambient conditions toprovide an equimolar solutions of Compound 1 and S-camphor sulfonicacid. The solution was allowed to stir for 48 hours at ambientconditions; crystallization occurred during that time. Followingfiltration and washing with isopropyl alcohol, the product was driedunder vacuum at 40° C. which provided approximately 75 mg of crystallinematerial.

Example 6: Characterization of the S-Camsylate Salt of Compound 1,Polymorph Form a and the Maleate Salt of Compound 1, Polymorph Form B byPowder X-Ray Diffraction (PXRD)

The powder X-ray diffraction patterns, as shown in FIGS. 3, 4, 9, and10, were determined using a Bruker-AXS Ltd. D4 powder X-raydiffractometer fitted with an automatic sample changer, a theta-thetagoniometer, automatic beam divergence slit, and a PSD Vantec-1 detector.The sample was prepared for analysis by mounting on a low backgroundcavity silicon wafer specimen mount. The specimen was rotated whilebeing irradiated with copper K-alpha₁ X-rays (wavelength=1.5406 Å) withthe X-ray tube operated at 40 kV/35 mA. The analyses were performed withthe goniometer running in continuous mode set for a 0.2 second count per0.018° step over a two theta range of 2° to 55°. Peaks were alignedagainst those of the calculated simulated powder pattern.

Example 7: Characterization of the Maleate Salt of Compound 1, PolymorphForm A by Powder X-Ray Diffraction (PXRD)

The powder X-ray diffraction (PXRD) pattern measurement, as shown inFIG. 1, was carried out on a Bruker D5000 diffractometer using copperradiation (CuKα, wavelength: 1.54056 Å). The tube voltage and amperagewere set to 40 kV and 40 mA, respectively. The divergence and scatteringslits were set at 1 mm, and the receiving slit was set at 0.6 mm.Diffracted radiation was detected by a Kevex PSI detector. A theta-twotheta continuous scan at 2.4 degrees/min (1 second/0.04 degree step)from 3.0 to 40 degrees 2θ was used. An alumina standard was analyzed tocheck the instrument alignment. Samples were prepared by placing them ina quartz holder.

Example 8: Characterization of the S-Camsylate Salt of Compound 1,Polymorph Form B by Powder X-Ray Diffraction (PXRD)

The powder X-ray diffraction pattern, as shown in FIG. 15, was obtainedusing a Bruker AXS Ltd. D8 Advance powder X-ray diffractometer fittedwith Gobel mirror optics, a single sample heating stage and a positionsensitive detector (PSD). Each specimen was irradiated with copperK-alpha₁ X-rays (wavelength=1.5406 Å) with the X-ray tube operated at 40kV/40 mA. Analysis was performed with the goniometer running incontinuous scan mode set for a 0.2 second count per 0.014° step over arange of 3° to 35° 2θ. Measurement was performed at 150° C. with thetemperature controlled using an Ansyco sycos-H-HOT temperaturecontroller.

Example 9: Characterization of the Maleate Salt of Compound 1, PolymorphForm B by Solid State Nuclear Magnetic Resonance (SSNMR)

Spectra were collected at ambient temperature and pressure on aBruker-Biospin 4 mm BL CPMAS probe positioned into a wide-boreBruker-Biospin DSX 500 MHz (¹H frequency) NMR spectrometer. The packedrotor was oriented at the magic angle and spun at 15.0 kHz. The ¹³Csolid state spectrum, as shown in FIG. 5, was collected using a protondecoupled cross-polarization magic angle spinning (CPMAS). Thecross-polarization contact time was set to 2.0 ms. A proton decouplingfield of approximately 85 kHz was applied. 4096 scans were collectedwith a 14 second recycle delay. The carbon spectrum was referenced usingan external standard of crystalline adamantane, setting its upfieldresonance to 29.5 ppm. The ¹⁹F solid state spectrum, as shown in FIG. 6,was collected using a proton decoupled magic angle spinning experiment(MAS). A proton decoupling field of approximately 85 kHz was applied.128 scans were collected with recycle delay of 140 seconds. The fluorinespectrum was referenced using an external standard of trifluoroaceticacid (50% V/V in H₂O), setting its resonance to −76.54 ppm.

Example 10: Characterization of the S-Camsylate Salt of Compound 1,Polymorph Form a by Solid State Nuclear Magnetic Resonance (SSNMR)

Approximately 80 mg of sample were tightly packed into a 4 mm ZrO₂rotor. Spectra were collected at ambient temperature and pressure on aBruker-Biospin 4 mm BL CPMAS probe positioned into a wide-boreBruker-Biospin DSX 500 MHz (¹H frequency) NMR spectrometer. The packedrotor was oriented at the magic angle and spun at 15.0 kHz. The ¹³Csolid state spectrum, as shown in FIG. 11, was collected using a protondecoupled cross-polarization magic angle spinning (CPMAS). Thecross-polarization contact time was set to 2.0 ms. A proton decouplingfield of approximately 85 kHz was applied. 2048 scans were collectedwith a 6 second recycle delay. The carbon spectrum was referenced usingan external standard of crystalline adamantane, setting its upfieldresonance to 29.5 ppm. The ¹⁹F solid state spectrum, as shown in FIG.12, was collected using a proton decoupled magic angle spinningexperiment (MAS). A proton decoupling field of approximately 85 kHz wasapplied. 256 scans were collected with recycle delay of 28 seconds. Thefluorine spectrum was referenced using an external standard oftrifluoroacetic acid (50% V/V in H₂O), setting its resonance to −76.54ppm.

Example 11: Characterization of Polymorphs of Compound 1 by DifferentialScanning Calorimetry (DSC)

Differential Scanning Calorimetry of various polymorphs, as shown inFIGS. 2, 7, 13, and 24-27, was performed using a TA Instruments Q1000 ora Mettler Instruments DSC822. Samples (1 to 2 mg) were heated in acrimped aluminum pans from 20° C. at 10° C. per minute with a nitrogengas purge, up to as much as about 320° C.

Example 12: Characterization of Polymorphs of Compound 1 by DynamicVapor Sorption (DVS)

Hygroscopicity, as shown in FIGS. 8 and 14, was measured using anAutomated Sorption Analyser Model DVS-1, manufactured by SurfaceMeasurements Systems Ltd. UK. Solid (20-25 mg) was exposed to acontrolled relative humidity (% RH) and temperature environment (30°C.), and the weight change was recorded over time. The humidity wasstepped from 0 to 90% RH in 15% RH intervals. A rate of sorption of0.0005%/min averaged over 10 min was achieved at each humidity prior toexposure to the next humidity in the method.

Example 13: Preparation of a Solid Dosage Form of the S-camsylate Saltof Compound 1, Polymorph Form A

The S-camsylate salt polymorph Form A of Compound 1 was formulated intoimmediate release tablets. The formulated composition contained thefollowing components:

Quantity/unit: Component: (%) S-camsylate polymorph Form A 17.18Polymorph of Compound 1 Microcrystalline cellulose 52.55 Dicalciumphosphate anhydrous 26.27 Sodium Starch Glycolate (Type A) 3 MagnesiumStearate 1 Total: 100The formulated composition was characterized by the PXRD pattern shownin FIG. 16.

The same or similar formulation as above can be made using the maleatesalt polymorphs, such that the same or a similar amount of free basedrug concentration is maintained in the maleate salt formulation as inthe formulation above.

Example 14: Physical Stability of Maleate Polymorph Form B

A PXRD pattern for maleate polymorph Form B was measured at: 1) aninitial time point and 2): two weeks after storage at 70° C. with 75%relative humidity (RH). The PXRD pattern of maleate polymorph Form B didnot change significantly after two weeks of storage at 70° C. with 75%relative humidity. This demonstrates that maleate polymorph Form Bexists in a physically stable form.

Example 15: Physical Stability of S-Camsylate Polymorph Form A

A PXRD pattern for S-camsylate polymorph Form A was measured at: 1) aninitial time point and 2): two weeks after storage at 70° C. with 75%relative humidity (RH). The PXRD pattern of S-camsylate polymorph Form Adid not change significantly after two weeks of storage at 70° C. with75% relative humidity. This demonstrates that S-camsylate polymorph FormA exists in a physically stable form.

While the invention has been illustrated by reference to specific andpreferred embodiments, those skilled in the art will recognize thatvariations and modifications may be made through routine experimentationand practice of the invention. Thus, the invention is intended not to belimited by the foregoing description, but to be defined by the appendedclaims and their equivalents.

Example 16: Preparation of an S-Camsylate Salt of Compound 1,S-Camsylate Polymorph Form C

A slurry of S-Camsylate polymorph Form A (1 g) was prepared in isopropylalcohol:water (10 mL; 40:60% v/v). The slurry was heated to 70° C. overa 10 minute period to obtain a solution. The solution was cooled to 25°C. to obtain a supersaturated solution. Isopropyl alcohol:water (25 mL;10:90% v/v) and water (30 mL) were added. The resultant supersaturatedsolution was transferred to a rotary evaporator and the solvent removedunder vacuum (50 mbar) at 70° C. A precipitate was formed and isolated(0.6 g).

Example 17: Preparation of a 1R:1S-Camsylate Salt

A slurry of Compound 1 (1.5 g) was prepared in isopropyl alcohol:water(25 mL; 40:60% v/v). R-camphor sulfonic acid (0.65 g) and S-camphorsulfonic acid (0.65 g) were added, as a solution, in water (1.5 mL). Theslurry was heated to 70° C. over a 10 minute period. The resultantsolution was cooled to 0° C. over a 10 minute period. Solid crystallizedafter holding this solution at a temperature of 0° C. for one hour. Thisresulted in the formation of a slurry. This slurry was granulated for atotal of 36 hours.

The crystals were filtered and washed with water and then driedovernight at 50° C. providing a pale yellow powder (1.9 g).

Example 18: Preparation of a 1R:9S-Camsylate Salt

A slurry of Compound 1 (1.5 g) was prepared in isopropyl alcohol:water(25 mL; 40:60% v/v). R-camphor sulfonic acid (0.13 g) and S-camphorsulfonic acid (1.17 g) were added, as a solution, in water (1.5 mL). Theslurry was heated to 70° C. over a 10 minute period. The resultantsolution was cooled to 10° C. over a 10 minute period.

Solid crystallized after holding this solution at a temperature of 10°C. for one hour. This resulted in the formation of a slurry. This slurrywas granulated for a total of 48 hours.

The crystals were filtered and washed with water and then driedovernight at 50° C. providing a pale yellow powder.

Example 19: Preparation of a 1R:3S-Camsylate Salt

A slurry of Compound 1 (1.5 g) was prepared in isopropyl alcohol:water(25 mL; 40:60% v/v). R-camphor sulfonic acid (0.325 g) and S-camphorsulfonic acid (0.975 g) were added, as a solution, in water (1.5 mL).The slurry was heated to 70° C. over a 10 minute period. The resultantsolution was cooled to 10° C. over a 10 minute period. Solidcrystallized after holding this solution at a temperature of 10° C. Thisresulted in the formation of a slurry. This slurry was granulated for atotal of 4 hours. The crystals were filtered and washed with water andthen dried overnight at 50° C. providing a pale yellow powder.

Example 20: Preparation of a 1R:7S-Camsylate Salt

A slurry of Compound 1 (1.5 g) was prepared in isopropyl alcohol:water(25 mL; 40:60% v/v). R-camphor sulfonic acid (0.16 g) and S-camphorsulfonic acid (1.14 g) were added, as a solution, in water (1.5 mL). Theslurry was heated to 70° C. over a 10 minute period. The resultantsolution was cooled to 10° C. over a 10 minute period. Solidcrystallized after holding this solution at a temperature of 10° C. Thisresulted in the formation of a slurry. This slurry was granulated for atotal of 4 hours. The crystals were filtered and washed with water andthen dried overnight at 50° C. providing a pale yellow powder.

Example 21: Preparation of an R-Camsylate Salt of Compound 1,R-Camsylate Polymorph Form A

A slurry of Compound 1 (1.5 g) was prepared in isopropyl alcohol:water(25 mL; 40:60% v/v). R-camphor sulfonic acid (1.3 g) was added, as asolution, in water (1.5 mL). The slurry was heated to 70° C. over a 10minute period. The resultant solution was cooled to 10° C. over a 10minute period. Solid crystallized after holding this solution at atemperature of 10° C. This resulted in the formation of a slurry. Thisslurry was granulated for a total of 4 hours. The crystals were filteredand washed with water and then dried overnight at 50° C. providing apale yellow powder.

Example 22: Characterization of the S-Camsylate Salt of Compound 1,Polymorph Form C, the 1R:1S-Camsylate Salt, the 1R:9S-Camsylate Salt,the 1R:3S-Camsylate Salt, the 1R:7S-Camsylate Salt, and the R-CamsylateSalt of Compound 1, R-Camsylate Polymorph Form A by Powder X-RayDiffraction (PXRD)

The powder X-ray diffraction patterns, as shown in FIGS. 18-23, weredetermined using a Bruker-AXS Ltd. D4 powder X-ray diffractometer fittedwith an automatic sample changer, a theta-theta goniometer, automaticbeam divergence slit, and a PSD Vantec-1 detector. The sample wasprepared for analysis by mounting on a low background cavity siliconwafer specimen mount. The specimen was rotated while being irradiatedwith copper K-alpha₁ X-rays (wavelength=1.5406 Ångstroms) with the X-raytube operated at 40 kV/35 mA. The analyses were performed with thegoniometer running in continuous mode set for a 0.2 second count per0.0180 step over a two theta range of 20 to 55°. Peaks were alignedagainst those of the calculated simulated powder pattern whereavailable. Alternatively, the peaks were aligned using an internalreference material, such as silicon or corundum (Al₂O₃), mixed with thepowder sample prior to analysis.

Example 23: Characterization of the S-Camsylate Salt of Compound 1,Polymorph Form C, by Solid State Nuclear Magnetic Resonance (SSNMR)

Approximately 80 mg of each sample was tightly packed into a 4 mm ZrO₂rotor. Spectra were collected at ambient conditions on a Bruker-Biospin4 mm BL CPMAS probe positioned into a wide-bore Bruker-Biospin DSX 500MHz (¹H frequency) NMR spectrometer. The packed rotor was oriented atthe magic angle and spun at 15.0 kHz. The ¹³C solid state spectrum, asshown in FIG. 28, was collected using a proton decoupledcross-polarization magic angle spinning (CPMAS) experiment. Thecross-polarization contact time was set to 2.0 ms. A proton decouplingfield of approximately 85 kHz was applied during acquisition. A minimumof 2048 scans were collected with a 7 second recycle delay. The carbonspectra were referenced using an external standard of crystallineadamantane, setting its upfield resonance to 29.5 ppm. The ¹⁹F solidstate spectrum, as shown in FIG. 29, was collected using a protondecoupled magic angle spinning (MAS) experiment. A proton decouplingfield of approximately 85 kHz was applied during acquisition. A minimumof 128 scans were collected with a recycle delay of approximately 30seconds. The fluorine spectrum was referenced using an external standardof trifluoroacetic acid (50% V/V in H₂O), setting its resonance to−76.54 ppm.

Example 24: Characterization of the 1R:1S-Camsylate Salt and the1R:9S-Camsylate Salt by Solid State Nuclear Magnetic Resonance (SSNMR)

Approximately 80 mg of each sample was tightly packed into a 4 mm ZrO₂rotor. Spectra were collected at ambient conditions on a Bruker-Biospin4 mm BL CPMAS probe positioned into a wide-bore Bruker-Biospin DSX 500MHz (¹H frequency) NMR spectrometer. The packed rotor was oriented atthe magic angle and spun at 15.0 kHz. The ¹³C solid state spectra, asshown in FIGS. 30 and 32, were collected using a proton decoupledcross-polarization magic angle spinning (CPMAS) experiment. Thecross-polarization contact time was set to 2.0 ms. A proton decouplingfield of approximately 85 kHz was applied during acquisition. A minimumof 2048 scans were collected with a 6 second recycle delay. The carbonspectra were referenced using an external standard of crystallineadamantane, setting its upfield resonance to 29.5 ppm.

The ¹⁹F solid state spectra, as shown in FIGS. 31 and 33, were collectedusing a proton decoupled magic angle spinning (MAS) experiment. A protondecoupling field of approximately 85 kHz was applied during acquisition.A minimum of 128 scans were collected with a recycle delay ofapproximately 30 seconds. The fluorine spectra were referenced using anexternal standard of trifluoroacetic acid (50% V/V in H₂O), setting itsresonance to −76.54 ppm.

Example 25: Characterization of Salts and Polymorphs of Compound 1 byFourier Transform-Infrared Spectroscopy (FT-IR)

The IR spectra were acquired using a ThermoNicolet Nexus FTIRspectrometer equipped with a ‘DurasampIIR’ single reflection ATRaccessory (diamond surface on zinc selenide substrate) and d-TGS KBrdetector. The spectra were collected at 2 cm⁻¹ resolution and aco-addition of 512 scans. Happ-Genzel apodization was used. Because theFT-IR spectra were recorded using single reflection ATR, no samplepreparation was required. Using ATR FT-IR will typically cause therelative intensities of infrared bands to differ from those seen in atransmission FT-IR spectrum using KBr disc or nujol mull samplepreparations. Due to the nature of ATR FT-IR, the bands at lowerwavenumber are typically more intense than those at higher wavenumber.Experimental error, unless otherwise noted, was ±2 cm⁻¹.

Example 26: Characterization of Salts and Polymorphs of Compound 1 byFourier Transform-Raman Spectroscopy (FT-Raman)

The Raman spectra were collected using a Bruker Vertex70 FT-IRspectrometer with RamII Raman module equipped with a 1064 nm NdYAG laserand LN-Germanium detector. All spectra were recorded using 2 cm⁻¹resolution and Blackman-Harris 4-term apodization. The laser power was250 mW and 1024 scans were co-added.

Example 27: Preparation of the Amorphous Form of the S-Camsylate Salt ofCompound 1

A solution of S-Camsylate polymorph Form A (150 mg) was prepared intBA:water (50 ml; 60:40% v/v) at room temperature. The solution wasfrozen by swirling on dry ice-acetone bath over a 4-5 minute period toobtain a thick frozen layer on the sides of the sample flask. Thecondenser of the lyophilizer was cooled to −100° C. and vacuum wasswitched on. Sample flask with frozen solution was quickly attached tothe port of a manifold or drying chamber. The vacuum was created byopening the vent to the chamber. The amorphous form of the S-camsylatesalt of Compound 1 was isolated after overnight drying at roomtemperature.

Example 28: Characterization of the Amorphous Form of the S-CamsylateSalt of Compound 1 by Powder X-ray Diffraction (PXRD)

The powder X-ray diffraction pattern was obtained using a Bruker-AXSLtd. D4 powder X-ray diffractometer fitted with an automatic samplechanger, a theta-theta goniometer, automatic beam divergence slit, and aLynxEye detector. The sample was prepared for analysis by mounting on alow background cavity silicon wafer specimen mount. The specimen wasrotated while being irradiated with copper K-alpha₁ X-rays(wavelength=1.5406 Ångstroms) with the X-ray tube operated at 40 kV/40mA. The analysis were performed with the goniometer running incontinuous mode set for a 0.3 second count per 0.0200 step over a twotheta range of 3° to 400. The PXRD diffractogram, as shown in FIG. 34,exhibits a broad peak having a base that extends from about 5° 2θ toabout 40° 2θ.

Example 29: Characterization of the Amorphous Form of the S-CamsylateSalt of Compound 1 by Solid State Nuclear Magnetic Resonance (SSNMR)

Approximately 80 mg of sample was tightly packed into a 4 mm ZrO₂ rotor.Spectra were collected on a Bruker-Biospin 4 mm BL CPMAS probepositioned into a wide-bore Bruker-Biospin DSX 500 MHz (¹H frequency)NMR spectrometer. The packed rotor was oriented at the magic angle andspun at 15.0 kHz. The rotor was cooled with a direct stream of nitrogenhaving an output temperature of 0° C. The ¹³C solid state spectrum, asshown in FIG. 35, was collected using a proton decoupledcross-polarization magic angle spinning (CPMAS) experiment. Thecross-polarization contact time was set to 2.0 ms. A proton decouplingfield of approximately 85 kHz was applied during acquisition. 10240scans were collected with a 5.5 second recycle delay. The carbonspectrum was referenced using an external standard of crystallineadamantane, setting its upfield resonance to 29.5 ppm. The ¹⁹F solidstate spectrum, as shown in FIG. 36, was collected using a protondecoupled magic angle spinning (MAS) experiment. A proton decouplingfield of approximately 85 kHz was applied during acquisition. 512 scanswere collected with a recycle delay of 5.5 seconds. The fluorinespectrum was referenced using an external standard of trifluoroaceticacid (50% V/V in H₂O), setting its resonance to −76.54 ppm.

Example 30: Characterization of the Amorphous Form of the S-CamsylateSalt of Compound 1 by Raman Spectroscopy

Raman spectra were collected using a Nicolet NXR FT-Raman accessoryattached to the FT-IR bench. The spectrometer was equipped with a 1064nm Nd:YAG laser and a liquid nitrogen cooled Germanium detector. Priorto data acquisition, instrument performance and calibrationverifications were conducted using polystyrene. Samples were analyzed inglass NMR tubes that were spun during spectral collection. The spectrawere collected using 0.5 W of laser power and 100 co-added scans. Thecollection range was 3700-300 cm⁻¹. All spectra were recorded using 4cm⁻¹ resolution and Happ-Genzel apodization.

Two separate spectra were recorded for each sample, which weresubsequently averaged and intensity normalized prior to peak picking.Peaks were manually identified using the Thermo Nicolet Omnic 7.3asoftware. Peak position was picked at the peak maximum, and peaks wereonly identified as such, if there was a slope on each side; shoulders onpeaks were not included. The peak position was rounded to the nearestwhole number.

Example 31: Characterization of the Amorphous Form of the S-CamsylateSalt of Compound 1 by Differential Scanning Calorimetry (DSC)

Differential Scanning Calorimetry (DSC), as shown in FIG. 38, wasperformed with a TA DSC (Q1000) Samples of approximately 5 mg wereweighed into Perkin Elmer hermetic aluminum pans (40 μl). Glasstransition temperature (Tg) measurement was conducted at 2° C./minuteheating rate with 1° C. amplitude and 100 seconds frequency in the −50to 200° C. The nitrogen purge was 50 mL/minute unless otherwise noted.The temperature was calibrated using indium.

The Tg of 156.5° C. obtained is the midpoint of the step transition athalf height in the reversing signal. Tg can change as a function ofwater and or solvent content.

We claim:
 1. A crystalline camsylate salt of8-fluoro-2-{4-[(methylamino)methyl]phenyl}-1,3,4,5-tetrahydro-6H-azepino[5,4,3-cd]indol-6-one,wherein the salt has a powder X-ray diffraction pattern comprising twoor more peaks at diffraction angle (2θ) selected from the groupconsisting of 12.2±0.2, 13.8±0.2, 18.3±0.2, 22.5±0.2, and 25.4±0.2,wherein said powder x-ray diffraction pattern is obtained using copperk-alpha₁ x-rays at a wave length of 1.5406 Ångstroms.
 2. The salt ofclaim 1, wherein the salt has a solid state NMR spectrum comprising 2 ormore ¹³C chemical shifts selected from the group consisting of213.4±0.2, 171.8±0.2, and 17.3±0.2 ppm.
 3. The salt of claim 1, whereinthe salt has a solid state NMR spectrum comprising ¹⁹F chemical shiftsat −118.9±0.2 and −119.7±0.2 ppm.
 4. The salt of claim 1, wherein theDSC thermogram of the salt comprises an endotherm onset at 303.2° C. 5.The salt of claim 1, wherein the camsylate comprises S-camsylate.
 6. Thesalt of claim 1, wherein the camsylate comprises R-camsylate.
 7. Apharmaceutical composition comprising the salt of claim 1 and apharmaceutically acceptable carrier.